Biological Use Authorization (BUA) Application FAQs

These frequently asked questions (FAQs) are designed to help you complete the BUA application. The below FAQs correspond to the new/renewal BUA application. The FAQs also pertain to the Change to BUA application, but the numbering of the referenced questions may differ.

rDNA refers to recombinant or synthetic DNA or RNA.

Refer to Biological Research Approval for information about the review process.

General Project Information

When do I submit a BUA change application? Should I submit it for every new cell line, gene insert, or microorganism? Should personnel changes be listed on a BUA change application?

Submit a BUA change application to add new research involving biohazards and new locations where biohazards will be handled. A BUA change is required to add new microorganisms, new viral vectors and/or gene inserts, new modified cell lines, new rDNA research, and any new biohazardous agents in animals. New cells lines and animal species may also require a BUA update. A BUA update is not required to add or remove personnel from your lab; personnel information is gathered only on new and renewal applications. If you are not sure if a BUA change is required, reach out to us at

Do I submit an IACUC protocol or a BUA first? What does the timeline look like for approval of biohazardous agents in animals?

Use of biohazardous agents in animals (including rDNA) requires BUA and IACUC approval. Because both the BUA and IACUC protocols need to be reviewed simultaneously for congruence, you should submit your BUA application or BUA change concurrently with the submission of the IACUC protocol or amendment. Submit the BUA as soon as possible to allow time for Institutional Biosafety Committee (IBC) review. Refer to the IBC meeting submission deadlines. For IACUC protocol renewals, it is recommended that you submit the corresponding BUA application two months prior to the IACUC protocol expiration to allow time for IBC review.

The IACUC protocol cannot be approved until the BUA is approved. If your BUA is approved by EH&S and the IBC, it will be held until the IACUC protocol reaches committee review. If you have questions about the approval timeline for your BUA submission, contact your assigned Biosafety Officer or the IBC coordinator at

I am a postdoctoral fellow/lab manager. Can I serve as Principal Investigator (PI) for submission of the BUA application?

No, it is the PI who is solely responsible for the accuracy of the BUA application and compliance with the applicable regulations. The lab contact section of the application is meant only for contact purposes, to request additional information, and to schedule laboratory inspections. Refer to the FAQ for the Statement of Responsibility.

Once I submit my BUA application, can I start my work right away?

Even though the National Institutes of Health (NIH) categorize certain types of experiments that can be initiated simultaneously with submission of the BUA application, institutional policy requires that you wait for approval from the IBC and/or EH&S.

  • For reviews covered under NIH Guidelines Sections III-A through III-C, contact EH&S for specific instructions.
  • For reviews covered under NIH Guidelines Section III-D, an IBC member and an EH&S biosafety officer will be assigned to your project as primary reviewers and will work with you to obtain approval. All IBC members will also have the opportunity to review your BUA application. You can proceed with your Section III-D research project after you receive your BUA letter.
  • For reviews covered under NIH Guidelines Section III-E, an EH&S biosafety officer will review the application and provide a preliminary approval on behalf of the IBC. The application is then reviewed at the next convened IBC meeting for final approval. You can proceed with your Section III-E research project after you receive your BUA letter.
  • Reviews falling under NIH Guidelines Section III-F are exempt from the NIH Guidelines (e.g., work with E. coli K-12 and its derivatives). However, to ensure compliance, the IBC recommends registering the research by submitting a BUA application to EH&S.
  • For research not subject to the NIH Guidelines including biohazardous agents that are not recombinant (e.g., wildtype microorganisms, human cell lines), an EH&S biosafety officer will be assigned to your project and will work with you to obtain approval. You can proceed with your research project after you receive your BUA letter.

The IBC review process includes a biosafety laboratory inspection of locations where biohazards are used that is conducted by an EH&S biosafety officer to evaluate lab space, containment practices, training, work procedures, and personal protective equipment (PPE). Once your research has been approved, you will receive a BUA letter from the IBC. If your research is not approved, an EH&S biosafety officer will work with you to address outstanding issues; approval is issued upon resolution.

How do I schedule a biosafety lab inspection? Are the inspection checklists available online? How do I respond to a biosafety lab inspection, and is there any timeline on when I should respond?

After you submit a BUA application, a biosafety officer will contact you regarding the biosafety lab inspection. Biosafety inspection checklists are on the UW EHS Biological Research Safety website in the Biosafety laboratory inspections section and linked directly here: BSL-1 inspection checklist, BSL-2 inspection checklist, and BSL-2 with BSL-3 practices checklist.

After an inspection, your assigned biosafety officer will send a report of findings and actions needed to address each finding. You can respond to biosafety inspection findings in the EHSA portal. For any questions or concerns regarding biosafety inspection findings or using EHSA to respond, please reach out to your assigned biosafety officer. Our Biological Inspections Guide shows you how to use the EHSA portal.

Submit responses to findings as soon as they are resolved. We aim to complete the biosafety lab inspection and response within 30 days or less, but some items may require more time to address. If you expect delays in your response, communicate that to your biosafety officer. BUA approval is contingent on completion of the biosafety lab inspection.

When should I provide an IACUC (Institutional Animal Care and Use Committee) protocol number?

Only one IACUC protocol can be associated with a single BUA, and the PI must be the same for the IACUC and the BUA. Provide an IACUC protocol number only if that IACUC protocol includes research involving biohazards administered to animals as described in the BUA application. If your research involves any work with transgenic animals or administration of biohazardous agents to animals, including rDNA, provide the IACUC protocol number.

If you collaborate with another PI or are involved with animal research covered under another PI’s IACUC protocol, do not list that IACUC number or the associated research in your BUA.

My project involves work with animals. Do I need approval for all my in vitro and in vivo work or just the work tied to my animal protocol?

You need approval for all your in vitro and in vivo work involving biohazards on a BUA. Each animal protocol involving biohazardous agents must have a separate BUA. If you have work with biohazardous agents unrelated to your animal protocol(s), then you may either:

  1. Include in vitro work on the BUA application associated with one of your animal protocols; or
  2. Submit a separate BUA application for the in vitro work.

What if I plan to have multiple IACUC protocols involving biohazardous agents in animals?

You will need to submit separate BUA applications for each IACUC protocol. Only one IACUC protocol can be associated with a single BUA. This also helps to ensure synchronized expirations dates for the associated BUA and IACUC.

I have not yet submitted my animal protocol to the Office of Animal Welfare (OAW). Can I still include the animal work on this BUA application?

Yes, you may include the animal work on the BUA application, but the risk assessment and IBC review cannot be completed until the IACUC protocol is submitted and reviewed by EH&S. It is best to submit the BUA application at the same time you submit your animal protocol. If you are not planning to start the animal work right away, you may want to submit your initial BUA application for in vitro work only. Later, you can submit a BUA change to add the in vivo work when you submit the animal protocol. IBC approval always precedes IACUC approval.

When should I provide a Human Subjects Division number?

Provide a Human Subjects Division number only when applicable to the research proposed in the BUA application; otherwise, leave the field blank. Specifically, if your research involves use of human source materials that require Institutional Review Board (IRB) approval or administration of biohazardous materials to human subjects (e.g., recombinant DNA vaccine), you must provide your Human Subjects Division number.

Research Description

Q1. What information should I provide in Question 1?

Provide only a brief and general description of the overall goals of the proposed research (one or two sentences). This information is used to help assign your project to reviewers with relevant expertise and to help them understand the background of your research. Provide your description in laymen's terms so that it can be understood by all members of the IBC, which includes professional staff, faculty from a variety of disciplines, and members of the public. The review process does not assess the scientific merit of the proposed research.

Q2. What information should I provide in Question 2?

Provide a brief but comprehensive description of the actual experimental procedures and biohazardous agents including rDNA that will be used for the proposed research. You are welcome to use a narrative or other formats as you wish, but all research involving biohazards needs to be accounted for in this question. You are strongly encouraged to use general terms for common laboratory procedures such as the following:

  • Engineering of plasmids in laboratory strain E. coli
  • Culture of human cell lines
  • Sorting human cells by flow cytometry

However, also be specific about which biohazards will be associated with which procedures. Some examples include the following.

  • Breeding mice containing the transgene for GFP
  • Transducing human cell lines with an adenoviral vector expressing the gene MYC
  • Working with a lentiviral vector expressing a GFP reporter gene

Be sure to specifically mention procedures that preclude the use of physical containment that would otherwise be required (e.g., stereotactic injections if Risk Group 2 viral vectors to animals, imaging of animals exposed to human cells when the imaging equipment cannot fit inside a biosafety cabinet). You may want to complete this question after you have filled out the rest of the application to ensure that all relevant procedures and biohazardous materials/agents are included. Be aware that your application may be returned to you as incomplete if this question is not consistent with the rest of your application.

Q2. What information should I provide about my work involving rDNA in Question 2?

If your research includes rDNA from any portion of one or a few genes, explicitly list the genes as well as the context in which the DNA or RNA will be used. If your research involves a large number of genes, then describe the functional categories of the proteins encoded by these genes, such as 'cytokine receptors' or 'cell signaling', as well as the context in which they will be used. If your research includes the discovery of new genes that will then be used in your research, describe the functional properties of the proteins encoded by the target genes (if known), how they will be identified (using general terms), and what you will do with them once they are identified.

You MUST use RefSeq gene names when listing specific genes. This is essential for reviewers to carry out a risk assessment of your proposed research. Be aware that your application may be returned to you as incomplete if you do not use RefSeq gene names. Instructions for verifying correct RefSeq gene names are included in the FAQ for question 36B.

Q2. What information should I not include in Question 2?

Do not provide experimental protocols or details such as dosing regimens or specific concentrations. Do not cut and paste procedures from animal protocols, grant proposals, or standard operating procedures. Do not provide the scientific justification, rational, or hypotheses associated with the proposed research.

Q3. What information should I include in Question 3?

Please describe what elements of your research that you believe, in your professional opinion, pose the greatest biohazardous risk to laboratory personnel or the environment. Be aware that a formal review of your application may lead to the identification of other biohazards associated with the proposed research which, in the opinion of biosafety professionals or members of the IBC, may constitute an even greater hazard. If this is the case, you may be contacted for further discussion on the topic. Be aware that your application may be returned to you as incomplete if you fail to identify what you believe to be the greatest biohazard associated with your proposed research.

Culture of Primary Cells or Cell Lines

Q15-18. Throughout the Culture of Primary Cells or Cell Lines section, I am asked to list the 'type' and 'source' of my cells/cell lines. What do you mean by type and source of cells/cell lines?

Type of primary cells means human or animal primary cells taken directly from blood or living tissue (e.g., peripheral blood mononuclear cells, T cells, dendritic cells, etc.).

Type of cell lines means human or animal cell lines that may or may not be well characterized [e.g., human derived HEK293 cell lines, Epstein-Barr Virus (EBV)-transformed lymphoblastoid cell lines, etc.].

If your cell lines contain rDNA, be sure to include them in the Recombinant and Synthetic DNA and RNA (rDNA) section, even if they were engineered prior to the application.

Q19-22 I work with induced pluripotent stem cells (iPSCs) that are purchased from an outside vendor, repository, or colleague. I will not be generating them in my laboratory. Do I need to fill out the Recombinant and Synthetic DNA and RNA and the Gene Delivery Methods table for this work?

Regardless of whether you will generate iPSCs in your laboratory or use iPSCs that already exist, you must include information here with the method of generation and gene inserts and then fill out relevant questions of the Recombinant and Synthetic DNA and RNA section. Also describe your use of these cells in your answer to Question 2.

Bloodborne Pathogens

Q23. I work only with established human cell lines obtained from a commercial vendor (e.g., ATCC) which is a very reliable and authentic source. I don't think the Washington State BBP rule applies to me. Please confirm.

The Washington State BBP Rule applies to all work done with human blood, tissue, body fluids visibly contaminated with blood or other potentially infectious materials (OPIM), as well as human cells and cell lines. All human cell lines, whether established or from authentic sources, are covered under the Washington State BBP Rule due to their potential to harbor BBPs. For a list of OPIM and information about the BBP program, refer to EH&S Bloodborne Pathogens

Q23. What information should I provide if I work with human tissue, blood, or body fluids, or culture of human primary cells or cell lines?

If you answered YES to questions 11, 15, 16, 19 and/or 20 for work with human blood, tissue, or body fluids, culture of human primary cells or cell lines, or iPSCs, the Washington State BBP Rule applies. As part of the BBP program, you will need to submit a site-specific BBP Exposure Control Plan. Initial and annual BBP training is also required. For more information, refer to Bloodborne Pathogens.

Bacteria, Viruses, Yeasts, Fungi, Parasites and Prions

Wildtype Microorganisms

Q24-25. My project involves work with non-recombinant microorganisms as well as recombinant microorganisms. In which sections of the BUA application should I list them?

For work involving non-recombinant microorganisms, including wildtype and naturally occurring mutant species, complete Questions 24-25.

In the table, provide the genus and species of the microorganisms you will be using along with any strain information that could impact the risk assessment. If you are working with naturally occurring mutant species, also provide information about the naturally occurring mutations and their effects on the microorganism. For examples of non-recombinant microorganisms, refer to the next FAQ.

For work involving recombinant microorganisms, complete Questions 26-30. You will also need to complete all applicable questions of the Recombinant and Synthetic DNA and RNA section. For the definition and examples of recombinant microorganisms, refer to the next FAQ. 

Q24-30. Can you provide some examples of non-recombinant and recombinant microorganisms?

Non-recombinant microorganisms include:

  • Wildtype, naturally occurring species of bacteria, viruses, yeasts, fungi, parasites and prions (e.g., Pseudomonas aeruginosaBacillus cereusTreponema pallidum)
  • Naturally occurring mutant species of bacteria, viruses, yeasts, fungi, parasites, and prions (e.g., Bacillus Calmette-GuerinChlamydia trachomatis LGV strains)

Recombinant microorganisms include:

  • Bacteria, viruses, yeasts, fungi, or parasites whose genetic material has been altered using rDNA technology. Examples include Listeria monocytogenes expressing ovalbumin, Toxoplasma gondii luciferase expressing PRU-Luc-GFP type II strain, and gene cloning done in E. coli K12.

Recombinant Microorganisms

Q26. What is a recombinant microorganism?

Refer to the previous FAQ for wildtype microorganisms for a description of recombinant and wildtype microorganisms.

Q26. Should I list viral vectors in the Recombinant Microorganisms questions and table?

No, viral vectors can be listed and described in the Recombinant or Synthetic Nucleic Acids (rDNA) section starting at Question 35.

Viral vectors are used to deliver genes to cells. They can either add a gene (knock-in) to study the function of a particular gene or to delete a gene (knock-down) to study the effects of gene deletion or reduction.  All work with viral vectors is subject to the NIH Guidelines for Recombinant or Synthetic Nucleic Acids, and as such, all work with viral vectors must be approved at a convened Institutional Biosafety Committee (IBC) meeting.

Q30. What do you mean by "potential for recombinant infectious agents to be released or shed from animals or plants?"

This question aims to capture shedding. If your project involves potential release or shedding of recombinant infectious agents from animals or plants, mark YES to Question 30. In your answer, explain why or how recombinant infectious agents may be released or shed.

Select Agents

Q31-34. Select Agents

I work with an excluded or attenuated strain of a Select Agent. Can you provide some examples of the rationale you would like for why the strain is exempt?

Select Agent excluded and attenuated strain information can be found on the CDC Select Agents and Toxins Exclusions website.

Examples of the rationale for an excluded or an attenuated Select Agent include:

  • A/Vietnam/1203 (H5N1) PR8 is excluded from the Select Agent regulations under Section 121.3 (e). This strain contains segments from A/PR/8/34 (H1N1) and H5 and N1 from Highly Pathogenic Avian Influenza A/Vietnam/1203/2004 (H5N1). The polybasic cleavage site in HA has been mutated to render it less pathogenic.  
  • Venezuelan equine encephalitis (VEE) subtypes ID and IE are excluded from the Select Agent regulations under Section 73.4 (e) and 121.4 (e).
  • Venezuelan equine encephalitis virus (VEEV) strain TC-83 excluded from the Select Agent regulations under Section 73.4 (e) and 121.4 (e). The strain is a vaccine strain of VEE.
  • West African clade of Monkeypox virus is excluded from the Select Agent regulations under Section 73.3 (e). Clinically severe human disease associated with West African strains is rare, and this virus clade has not been associated with human mortality.
  • Ebola ΔVP30 is excluded from the Select Agent regulations under Section 73.3 (e). This virus lacks the gene encoding for the VP30 protein. Therefore, the virus is replication incompetent and does not form infectious progeny in wild-type cells and does not cause disease in animals.

Q33. I work with nucleic acids from select agents. Do I need to complete this section?

Yes, complete this section if you work with or propose to work with positive strand RNA forms of Select Agent viral genomes or nucleic acids encoding for the toxic form(s) of regulated Select Toxins.

Recombinant and Synthetic DNA and RNA (rDNA)

Q35. How do you define recombinant or synthetic nucleic acid (rDNA) molecules?

Recombinant and synthetic nucleic acid molecules or "rDNA" molecules, including those that are chemically or otherwise modified analogs of nucleotides (e.g., morpholinos) or both, are defined in the context of the NIH Guidelines as follows.

  1. Molecules that (1) are constructed by joining nucleic acid molecules and (2) can replicate in a living cell (i.e., recombinant nucleic acids).
  2. Nucleic acid molecules that are chemically or by other means synthesized or amplified, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules (i.e., synthetic nucleic acids).
  3. Molecules that result from the replication of those described in (1) or (2) above.

The above definition reflects the amendment to the NIH Guidelines and the definition of rDNA stipulated in UW Administrative Policy Statement 12.3.

Examples of rDNA molecules outside of living cells that fall under the above definition include the following.

  • Cloning DNA in bacterial plasmids
  • siRNA
  • Oligonucleotide primers
  • naked DNA from PCR or DNA sequencing
  • base-pair analogs
  • Note that naked DNA molecules in test-tubes are exempt from the NIH Guidelines; however, when in an organism, naked DNA molecules are not exempt.

All forms of rDNA are treated as biohazards regardless of whether the rDNA molecules are (a) purchased from a vendor (e.g., purchase a human cell line containing a plasmid), (b) constructed in your laboratory, or (c) used to transfect cells. All rDNA, irrespective of exemption from the NIH Guidelines, must be treated as a biohazard. Refer to the Biological Research Approval webpage for the IBC's working definition of a biohazard. Containment practices for work with rDNA in BSL-1 or higher laboratories are as stipulated in the UW Biosafety Manual. Refer to Biohazardous Waste for information on how to properly identify, package, decontaminate, and dispose of biohazardous waste, including rDNA waste. 

Q36-44. What levels of review are required for different sections of the NIH Guidelines? Can you help me identify sections of the NIH Guidelines that are applicable to my work?

PIs are required to identify the sections of the NIH Guidelines that apply to their work. If you misidentify the sections of the NIH Guidelines that apply, a biosafety officer will notify you and help you to determine the correct sections. The EH&S Biosafety training includes an introduction to the NIH Guidelines sections.

Experiments Covered by the NIH Guidelines describes the levels of review necessary for certain types of experiments involving rDNA. The six categories of review reflect the risks associated with the research, with more stringent review required for higher risk experiments.

Section III-F: Experiments covered under this section are exempt from the NIH Guidelines. This includes rDNA that is not in organisms or viruses, use of rDNA in certain exempt microorganisms (i.e., E.coli K-12 strains), and use of transgenic rodents requiring ABSL-1 containment. Refer to Section III-F of the NIH Guidelines for specifcs.

Section III-E: Experiments covered under this section include use of rDNA in Risk Group 1 microorganisms or formulated into synthetic or natural vehicles, and research with whole plants at biosafety level P1 (Plants-1). Examples include the following.

  • E. coli, non-pathogenic strains [that are not exempt under Section III-F]
  • Pseudomonas graminis, rhizobium tropici plant pathogens used in plants at P1
  • All rDNA experiments in plants (e.g., plants infected with recombinant Amycolata autotrophica, Sphaerophorus necrophorus)

Section III-D: Experiments covered under this section include use of rDNA in pathogenic microorganisms, animals, plants, viral vectors for gene transfer, gene transfer in Risk Group 2 and 3 agents, and restricted agents. Examples include the following.

  • All viral vectors
  • All rDNA experiments in whole animals (e.g., mice injected with plasmids, replication deficient lentivirus, gammaretrovirus, adeno-associated virus, etc.)
  • Research involving more than ten liters of rDNA culture
  • Research with influenza viruses containing genes or segments from 1918 H1N1, human H2N2 (1957-1968), and highly pathogenic avian influenza H5N1 strains

Section III-C: Experiments covered under this section involve the deliberate transfer of rDNA or DNA/RNA derived from rDNA, into one or more human research participants. For more information on research involving rDNA in humans see Clinical Trials.

Section III-B: Experiments covered under this section involve the deliberate formation of rDNA containing genes for the biosynthesis of toxin molecules lethal for vertebrates at an LD50 of less than 100 nanograms per kilogram body weight (e.g., cloning in E. coli K12 of DNA containing genes coding for the biosynthesis of toxic molecules such as botulinum toxins, tetanus toxin, diphtheria toxin, Shigella dysenteriae neurotoxin).

Section III-A: Experiments covered under this section involve the deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the ability to control disease agents in humans, animals or agriculture. Research covered under Section III-A requires approval from the NIH Director. For more information see Major Actions under Section III-A of the NIH Guidelines.

Q38.h. Why do I need to fill out an additional form for use of influenza viruses? How do I complete the form?

Due to the complex nature of influenza virus nomenclature, a method was needed to simplify the review of research with influenza viruses, especially Influenza A viruses. To simplify the process, EH&S compiled a list of currently approved influenza strains, their containment levels, and their starting NIH sections. 

To complete the form, you will need to identify all the influenza strains that your lab works with. If the strains are listed in Question 1 of the supplemental influenza form, then check Yes in the associated column A for the strain. If the strain is not listed in Question 1, then you will need to complete Question 2.

For each strain in Question 2, you will need to list the strain designation using the nomenclature at the top of the supplemental form. Then, you will need to include any strain or modification information that you have, the risk group, and indicate if the strain is recombinant.

Viral Vectors and Gene Delivery Methods Table

Q45. I work with infectious clones (e.g., cDNA clones), should I list them here?

Yes, work with cDNA clones needs to be listed in this table. Also describe your infectious clones and what you will be doing with them in Q38d.

Q45B. What are the different replication status options?

  • Replication defective and testing for RCV: Most viral vectors are engineered so that the viral nucleic acid packaged into the viral vector particle and transferred to recipient cells is insufficient to support replication of the vector and production of infectious progeny. Note that the NIH Guidelines (Section III-D) defines defective viruses as those containing less than 2/3 of the genome of any eukaryotic virus where all viruses from the same family are considered identical. Production of replication defective vectors requires complementation of essential elements in trans, often in the form of plasmids or packaging cells. Therefore, production of replication defective vectors often involves an inherent risk of reconstituting an intact or replication-competent virus (RCV), because all the genetic elements needed to produce infectious progeny may be found together in the producer cells. Some vector systems (e.g., third generation or later lentiviral vectors) incorporate a sufficiently high level of engineering safeguards to render the production of RCV almost impossible. However, for other commonly used viral vectors like amphotropic gammaretroviruses, RCV could persist undetected in transduced cells or animals indefinitely. This is a particular concern for vectors encoding oncogenes. Thus, in addition to the characteristics of the vector and the identity of the transgene, RCV testing may inform the appropriate biosafety levels for the use of viral vectors as well as cells, animals, and plants exposed to viral vectors. RCV testing varies depending on the viral vector, but common techniques include PCR tests for reconstitution of the genome, plaque assays for viral replication and spread, or detection of viral gene products (e.g., by ELISA). If RCV testing is performed, the results must be included with the BUA to be incorporated into the risk assessment.
  • Replication competent: Some applications require viral vectors capable of infecting cells and replicating. One example is the use of oncolytic vectors for cancer treatment. In this case, the genome may be modified so that viral replication occurs preferentially in certain cell types (e.g., transformed cells). Alternatively, or in addition, a transgene may be added to the genome in a way that does not interfere with replication. Information about the conditions under which the vector will replicate, any attenuation of pathogenicity or modification of tropism compared to the wild type virus, as well as the identity of the transgene are all important elements of risk assessment for these vectors.
  • Negative for replication competent virus (RCV): Selecting this indicates that you have had your viral vector or transduced cell line appropriately testing for the presence of RCV and the testing showed RCV is not present. If RCV testing has been performed on viral vectors, submit the RCV test results with the BUA application. More information about which viral vectors can be RCV tested and the appropriate tests is on our Viral Vectors for Gene Transfer webpage.
  • Other (describe): This is an option if needed for special circumstances.
  • Not applicable (non-viral): This applies for non-viral gene delivery methods such as plasmids, mRNA, siRNA, etc.

Q45C. I am asked to provide gene inserts or transgenes. Can I use gene names other than RefSeq gene names in column C of the Viral Vector and Gene Delivery Methods table?

No, gene names other than RefSeq gene names may not be used in this column. The IBC requires that you provide only RefSeq gene names. Some examples of RefSeq gene names include 'GFP', 'MYC', and 'COP4'. To verify that your gene names are correct follow the instructions below.

  1. Access the National Center for Biotechnology Information (NCBI) RefSeqGene website.
  2. Enter your gene name. Select 'Search' to search the RefSeq database.
  3. The search results for your gene will display. If your gene name appears as blue linked text, you have entered a correct RefSeq gene name.

If your gene is a synthetic construct and does not appear in RefSeq, provide a reference that describes the gene.

If your research involves more genes than can be easily listed in column C, list the functional categories or the categories of the encoded proteins in Question 48. 

Q45D. My project involves in vitro work using viral vectors. How should I complete column D of the Viral Vector and Gene Delivery Methods table?

If your project involves in vitro work, mark YES in column D. In your answer, specify the cell species/type in which the viral vector (or other rDNA) will be used and explain the activities. Refer to examples provided on the Viral Vectors and Gene Delivery Methods table.

Q45E. My project involves administering viral vectors to animals. How should I complete column E of the Viral Vector and Gene Delivery Methods table?

If your project involves in vivo work, mark YES in column D. In your answer, specify the animal species, method of administration, exactly what will be administered to the animal, and explain the activities. For example:

  • Third generation lentiviral vectors will be injected into mouse footpad
  • Human cells transduced with amphotropic gammaretroviral vectors will be xenografted onto rat forelimbs
  • mRNA vaccines will be administered to mice via IP injection

Replication Competent Viral Vectors and Transduced Cells

Q46. What is Question 46 asking for? Why do I need to provide this additional information?

Many forms of commonly used viral vectors have the potential to release or shed replication competent viral vectors from cells, animals, or plants, some of which may be capable of integrating into the genome of an infected cell. This can persist long after the cells are transduced (e.g., cells transduced with early generation retroviral vectors). Release of replication competent viral vectors has health and safety implications for those handling infected cells, animals, or plants. This is why the IBC recommends performing replication competent virus (RCV) testing or using later generation vectors with increased safety features (e.g., third generation lentiviral vectors).

Third Generation Lentiviral Vectors

Q47. What are third generation lentiviral vectors? Why do I need to provide additional information for these vectors?

The IBC has established a working definition of third generation lentiviral vectors. If vectors meet the definition of third generation, biocontainment may be lowered for use with oncogenic inserts. Refer to Third Generation Lentiviral Vectors for more information.

Third generation lentiviral vector systems improve on the safety of earlier lentiviral vector systems by splitting the necessary genes onto additional plasmids which renders them essentially incapable of generating replication-competent virus. The packaging system is split into two plasmids: one encoding Rev and one encoding Gag and Pol. Then Tat is eliminated from the third generation system through the addition of a chimeric 5' LTR fused to a heterologous promoter on the transfer plasmid. Thus, expression of the transgene from this promoter is no longer dependent on Tat transactivation. Finally, the third generation transfer plasmid can be packaged by either a second generation or third generation packaging system.

For more information on third generation lentiviral vectors and how they compare to second generation lentiviral vectors, refer to the Addgene Lentiviral Guide.

If you are unable to identify all four plasmids in your lentiviral vector system, the lentiviral vector will be listed as first/second/unknown generation on your BUA application.

Gene Inserts

Q48. I have already listed the names of my genes in the Viral Vector and Gene Delivery Methods table. Should I list them here again?

No, you do not need to list your genes again in Question 48. If your research involves more genes than can be easily listed in the table, list the functional categories or the categories of the encoded proteins in Question 48. Examples include cytokine receptors, cell signaling, cloning transcription factors, and HLA/MHC genes.

Oncogenes and Tumor Suppressor Genes

Q49-52. How do I complete Questions 49-52?

You are required to disclose the use of all oncogenes and tumor suppressor genes in this section. Follow these instructions to complete this section.

To search the two databases:

  1. Verify that you are using correct RefSeq gene names. Instructions for verifying correct RefSeq gene names are included in FAQ for Q45C.
  2. Using RefSeq gene names, search both databases for the gene and review the information:
    1. For the Cancer Gene Census database, search using the search box in the Census table, not the search box on the site's toolbar.
    2. For the Cancer Genetics Web database, you can find genes using the alphabetical list or use the search field.
  3. If any of your genes appear in the Cancer Gene Census database or are described as oncogenes or tumor suppressors in the Cancer Genetics Web database, mark YES to Question 49 and list the gene names.
  4. If your genes do not appear in the database but are well described in the scientific literature as oncogenes or tumor suppressors, mark YES to Question 50. In your answer, list the gene names, briefly describe why they are considered oncogenes, and provide references.
  5. If your genes do not appear in the database but you have reasons to believe they are oncogenes or tumor suppressors, mark YES to Question 51. In your answer, list the gene names, briefly describe why you consider them to be oncogenes or tumor suppressors, and provide references.

Q52. What is the point of Question 52? I believe my work with oncogenes or tumor-suppressor genes should not require an elevation in biocontainment, and I do not want to work at an elevated level of biocontainment. Can I petition the IBC to not increase the level of biocontainment for my work with these oncogenes or tumor-suppressor genes? 

If any of the genes you are using are identified as potential oncogenes or tumor suppressors in the previous questions, describe in Q52 whether you are overexpressing or dysregulating oncogenes or knocking down or silencing tumor suppressors. The IBC will consider this when setting the biosafety level for this work.

It is the IBC's policy that the addition of oncogenes or tumor suppressor genes to certain classes of viral vectors necessitates that work with these vectors is carried out at higher levels of biocontainment. This includes work with oncogenes or tumor suppressor genes in all classes of vectors based on retroviruses, adeno-associated viruses, and any other virus-based vectors that have a reasonable capacity for genomic integration.

Give a description of the type of expression for each oncogene or tumor suppressor in each vector system. This will help to determine the biocontainment level.

Examples include:

  1. I am overexpressing oncogene X.
  2. I am overexpressing tumor suppressor gene X.
  3. I am knocking down the expression of oncogene X.
  4. I am silencing tumor suppressor gene X.
  5. I am working with an oncogene but have deleted a functional domain known to be necessary for the oncogenic potential of the corresponding protein.
  6. I am working with an oncogene that appears in one of the oncogene databases, but there is scientific evidence that this gene is not a driver of oncogenesis.

Please provide references from scientific literature as applicable.

Transgenic Animals

Q53. What are the exemptions for transgenic rodents?

The following are exempt as stipulated in the NIH Guidelines, Section III-F.

  1. The purchase or transfer of transgenic rodents.
  2. Generation of BSL-1 transgenic rodents via breeding, with the following exceptions. Exceptions are as stipulated in NIH Guidelines, Appendix C.
    • Breeding of rodents that have a gene encoding more than fifty percent of an exogenous eukaryotic virus
    • Breeding of rodents in which the transgene is under the control of a gammaretroviral long terminal repeat (LTR)

Refer to the Animal Activities Table from NIH Office of Science Policy for a listing of animal experiments covered under the NIH Guidelines with references to the corresponding NIH Guidelines sections and biosafety levels.

The PI is responsible for assuring that the transgenes do not include gammaretroviral LTRs of more than one-half of a viral genome. There are a growing number of examples in which PIs have not reported this correctly. PIs are obligated to review the primary literature associated with the transgenic rodents that they wish to use in order to verify this.

Containment Requirements

Q54. I am asked to identify the containment requirements and biosafety levels of my laboratory. Can you explain the different biocontainment levels?

Each Principal Investigator (PI) is responsible for performing a risk assessment of their work to identify hazards. The risk assessment helps determine which biohazard containment and laboratory practices are appropriate. Refer to Biological Risk Assessment for more information, including Risk Groups and Biological Safety Levels and the associated references.

Biosafety levels are well described by in the CDC’s BMBL and in the NIH Guidelines:

  1. Laboratory biosafety levels (BSLs) refer to in vitro lab spaces or shared facilities where biohazardous agents are used. The four BSLs consist of combinations of facility design features, primary and secondar barriers, practices and procedures, and personal protective equipment (PPE).
  2. Animal facility biosafety levels (ABSLs) refer to labs or animal facility spaces where biohazardous agents and rDNA are used in animals or where transgenic animals are used. ABSLs correspond to increased levels of protection for research staff and the environment and are recommended as the minimal standard for activities involving laboratory animals exposed to biohazardous agents. The animal care and use environment can present unique hazards not found in standard research laboratories including bites and scratches, zoonotic disease, shedding of infectious agents, etc. Keeping this in mind, all additional animal facility standard operating procedures must be followed.
  3. Plant facility biosafety levels (BSL-Ps) refer to labs or greenhouse spaces where biohazardous agents are used with plants or where transgenic plants are used. There are specialized requirements for research involving transgenic plants or biohazardous agents used with plants. More information is available on the Plant Research Biosafety webpage.
  4. Arthropod biosafety levels (ACLs) refer to labs or facilities where arthropods are used. Arthropod facilities may be classified according to biohazardous agents used in arthropods or based on the risk or hazard of the arthropod itself. Contact EH&S Biological Safety if you are unsure what ACL applies to your lab.

If this question is completed incorrectly, a biosafety officer will notify you and help you to determine the correct biosafety levels for your laboratory.

Q55-57. I am asked to list biohazardous agents and activities. Since I already listed the activities in previous questions, do I need to list them here again? If so, what do I need to list?

Yes, please provide a summary of the biohazardous agents and the general activities that will be done in each room. Examples of what should be listed include:

  • Culture of human cell lines, E. coli, 3rd gen. lentiviral vectors, and Pseudomonas aeruginosa. Sequencing and analysis of clinical P. aeruginosa isolates.
  • Human and NHP cell infections with RG2 viruses. Culture of RG2 viruses and plasmid generation in E. coli K12 strains.
  • Aerosol exposure of mice to bacteria and administration of a recombinant DNA vaccine to mice.
  • Intranasal inoculation of mice to RG2 bacteria.
  • Cell sorting of human and non-human primate cells, imaging of mice exposed to lentiviral vectors, generation of human iPS cells.

Q56. I am asked to identify the biosafety levels of my animal facilities. I am not sure about the biocontainment levels for my animal work. Can you explain the different animal biocontainment levels?

Animal Biosafety Level (ABSL) classification refers to facilities, practices, and operational requirements applicable to work with animals that have been exposed to biohazardous agents. Animals exposed to biohazardous agents assigned to BSLs 1-4 are generally assigned into ABSLs 1-4, respectively. ABSLs correspond to increased levels of protection for research staff and the environment and are recommended as the minimal standard for activities involving laboratory animals exposed to biohazardous agents. Note that no ABSL-4 research is conducted at the UW. Refer to the Animal Activities Table from NIH Office of Science Policy for a listing of animal experiments covered under the NIH Guidelines with references to the corresponding NIH Guidelines sections and biosafety levels.

Practices and facility requirements for biosafety laboratories apply to research with animals as well. The animal care and use environment can present unique hazards not found in standard microbiological research laboratories (e.g., animals may generate aerosols, bite and scratch, be infected with a zoonotic agent, etc.) Keeping this in mind, all additional animal facility standard operating procedures must be followed.

If this question is completed incorrectly, a biosafety officer will notify you and help you to determine the correct animal biosafety levels for your laboratory.

Q57. I work in a laboratory room that is shared with other labs, but I don’t think it is a core facility. Should I list it in the core facilities section or in the in vitro/in vivo use section(s)?

List work with biohazards in shared laboratories in the in vitro and/or in vivo rooms questions. If you are unsure if a laboratory is a core facility, reach out to EH&S Biological Safety at or the biosafety officer assigned to review your BUA application. The biosafety safety office maintains a list of facilities considered core facilities. These sites are regularly inspected outside of the review cycle of individual projects. These facilities include the UW Greenhouse, Flow Cytometry cores, the stem cell core, MRI facilities, etc.

Other Hazards

Q83. I work with Select Agent toxins under the permissible limits. I described my work with them in Question 34.a. Do I need to relist them here and submit an SOP?

Yes, all toxins, regardless of Select Agent status, need to be listed in this question. SOPs and documented training are required for all toxins used in your laboratory. More information is available in EH&S Biological Toxin Safe Work Practices.


Q89. Why should I take the biosafety training?

As mandated by the NIH Guidelines, PIs must complete the EH&S Biosafety Training prior to their initiation of work with biohazardous agents and every three years thereafter. The training is also required for students, fellows, laboratory managers, research staff, and other staff with potential for exposure to biohazardous agents, including rDNA, at the UW.

The training covers roles and responsibilities when conducting research with biohazardous agents, including rDNA. The course also covers the review and approval process for biohazardous agents at the UW and the requirements governing their use (e.g., facilities, equipment, practices).

It is recommended that you take the training before completing this application. Be aware that your application cannot be approved until completion of training is verified. 

Q91. I only ship/transport Category B materials. Do I still need to take the Shipping Hazardous Materials Training?

If you are only shipping Category B materials, then no, the Shipping Hazardous Materials training is not required. However, if you work with more than just Category B materials or would like to learn more about the different classes of hazardous materials and their requirements, then the Shipping Hazardous Materials training is recommended.

 The Shipping Biological Substances Category B course meets the training requirements for shipping materials classified as a Biological substance, Category B, in the Department of Transportation (DOT) and International Air Transportation Association (IATA) regulations. The training includes instructions for packaging, labeling, and preparing shipping papers and is required to be taken every two years if you are actively shipping such materials.

Statement of Responsibility

Q. I am a postdoctoral fellow/lab manager and have prepared this BUA application, but I am not the PI of my laboratory. Can I sign the Statement of Responsibility on behalf of the PI?

NIH Guidelines, Section IV-B-7 states that the PI is the sole person responsible for the conditions set forth in the application, regardless of who prepared it. If the application was prepared by someone other than the PI, then the PI should take care to review the entire application in detail before signing the Statement of Responsibility.


Biological Safety Contact

(206) 221-7770