TAT PROTEIN GUIDELINES
The Institutional Biosafety Committee (IBC) has received numerous inquiries for investigators requesting to purchase/use Trans-Activating Transduction (TAT) proteins or other tags which promote protein entry into cells, in their research. Many investigators initially view TAT fusion protein expression vectors as just one of the many plasmids, which they may use in their laboratory and as such, submission to the IBC may be a surprising requirement. They may also view use of TAT fusion proteins outside the purview of the IBC.
The Committee's mandate is to review all research being conducted at the University of Chicago which involves the use of biohazards including -all- recombinant DNA, agents infectious to humans, animals or plants and other altered -(e.g. transgenic) oganisms. Expression of a TAT- fusion- protein, even in bacteria, is considered rDNA work and falls under the purview of the IBC as Risk Group 2. In addition, the TAT protein has potentially unique and unknown infectious qualities. As such, the use of the TAT protein, is categorized as biosafety level 2 (BSL2) and accidental spill procedures must be outlined in a protocol submission to the IBC. Protocols using TAT will be required to develop a safety manual and include it with the protocol submission.
TAT Protein Use
TAT Protein use may be pursued at the University of Chicago if the guidelines developed by the Institutional Biosafety Committee are followed.
Containment of work with TAT Protein
TAT Protein is categorized as Biosafety Level 2 (BSL2).
Laboratory Containment, Practice, and Technique:
- TAT Protein must be handled as a potentially hazardous material.
- Some proteins are more toxic and/or immunogenic and should be identified.
- Plastic backed absorbent lab paper should be used on all laboratory bench surfaces to absorb spills and splashes. All things that come in contact with TAT proteins should be regarded as contaminated.
- Biological safety cabinet (preferred) or designated space is recommended
- Avoid aerosol-generating activities or use appropriate safety equipment such as biological safety cabinets and sealed centrifuge tubes
Personal Protection required:
- Mouth pipetting is not allowed
- Lab coats
- Disposable latex, nitrile, or equivalent gloves
- Safety goggles
- Avoid direct contact with the skin, cuts, mucous membranes
- Wash well after working with TAT material.
In the event of a spill, while wearing gloves, labcoat, and safety glasses:
- Decontaminate work surfaces using a detergent with a protease enzyme (like Terg-A-Zyme) for 10-20 minutes
- Followed by washing with water and then wipe with a 70% ethyl alcohol solution.
- Deactivate and dispose of TAT solutions and cultures using standard autoclave.
- Dilute solutions can be deactivated using a 1:10 dilution of bleach (sodium hypochlorite solution) in a 1:1 mixture with the TAT solution, let sit for five to ten minutes. Dispose by sewer drain with copious amounts of water.
TAT Protein research approval
TAT Protein research must be approved by the IBC prior to its initiation. When any revision to an approved protocol is desired, an amendment must be filled with the IBC. The IBC reserves the right to approve exceptions to the above guidelines on a case by case basis. A protocol or an amendment to an existing protocol must be submitted to purchase, synthesize or express TAT proteins. The protocol or amendment must indicate:
- What peptide you are linking on to?
- What you are using as target cells?
- What are the harmful consequences, if any, when expressed?
Backus, B.D., Dowdy, S.F., Boschert, K.R., and Richards, T.L, Becker-Hapak, M. (2000). Safety Guidance for Laboratory Personnel Working with Trans-Activating Transduction (TAT) Protein Transduction Domains. American Chemical Society Journal of Chemical Health and Safety (submitted).