FAQ: Restricted Experiments

A request for approval to conduct a restricted experiment should be submitted by the entity’s Responsible Official and sent to the  Federal Select Agent Program (FSAP)through the electronic information system. The request will be reviewed by FSAP. We recommend that it not be submitted as part of an amendment as doing so may delay approval of other changes proposed in the amendment while the restricted experiment request is under review.

Submit a cover letter signed by the entity’s Responsible Official, on entity letterhead, requesting approval to conduct the restricted experiment, along with the following information (if applicable):

• Synopsis of the proposed restricted experiment and the intended objective(s).
• Description of the nucleic acid insert (complete sequence information is not required) and the predicted biological characteristics of the recombinant or synthetic product.
• Description of the cloning/expression vector.
• Identification and characteristics of the host organism used for molecular cloning.
• Description of the methods used for selection (e.g. recombinant or passive selection) to include all potential drug resistant traits (including intermediate variants).
• Address whether there are alternative drug resistance marker genes that could be used (i.e., to avoid the acquisition of drug resistance that could compromise the use of the drug to control disease agents in humans, veterinary medicine, or agriculture). Provide an explanation as to why these alternative marker genes are not being considered.
• Description of biosafety level including facility containment, equipment and special practices to be utilized for the proposed restricted experiment(s).
• Description of the mechanism and specificity of the drug resistance (antibiotic and antiviral resistance) being conferred to include any cross-resistance to other therapeutically useful antimicrobials.
• Estimated amount of toxin (recombinant or synthetic) to be produced.
• Synopsis of any planned animal or plant experiments or other relevant animal or plant work.
• Scientific references or supporting documentation, particularly with respect to the biosafety aspects of the proposed experimental product.

Yes. The request must describe all methods used for selection, including all intermediate vectors containing drug resistant markers or potential drug resistant traits. Refer to the Restricted Experiment Guidance Document for more information regarding submission of restricted experiment requests to the Federal Select Agent Program.

It depends on the circumstance. If the product of a restricted experiment is itself a select agent or toxin, the same approval to possess, use, and transfer the restricted experiment product is required under section 7 of the select agent regulations as for any other select agent or toxin. As to whether the product requires specific approval under section 13 of the regulations, it depends on when the product of the restricted experiment was created. If the product was created prior to December 4, 2012, the product of a restricted experiment does not require special approval for possession under section 13 of the select agent regulations. If the product was created on or after December 4, 2012, the possession is regulated under section 13 of the select agent regulations. For those regulated restricted experiment products, the entity must submit a written request to the Federal Select Agent Program asking permission to possess the product. If the possession of the product from a restricted experiment is approved, the entity must update the relevant information as an amendment to their select agent registration. In addition, the subsequent Principal Investigator (PI)’s work objective should be submitted for review, and the entity could expect that any approval would be subject to the same restrictions and biosafety requirements as were imposed by the initial “restricted experiment” approval. Please refer to the Restricted Experiment Guidance Document for more information regarding the regulation status of the possession of products from restricted experiments.

Yes. The entity must submit a written request to FSAP asking permission to conduct the experiment. If the request to perform the experiment is approved, the entity must update the relevant information as an amendment to their select agent registration. Only passive selection experiments conducted on or after December 4, 2012 are regulated under Section 13 of the select agent regulations. Please refer to the Restricted Experiment Guidance Document for more information regarding the regulation status to conduct restricted experiments using passive selection or utilizing recombinant DNA.

Depending on when the product was generated, the possession of a product of a restricted experiment may require pre-approval from FSAP before a transfer request is approved. Please refer to the Restricted Experiment Guidance document for a list of documentation required for review.

In addition, the APHIS/CDC Form 2, Request to Transfer Select Agents and Toxins, must be completed and submitted to FSAP prior to transferring any select agent(s) or any toxin(s) over the exclusion amount as indicated in section 3(d)(7). In Block 26, “characterization of agent” field, describe all drug resistance traits or recombinant DNA containing genes for the biosynthesis of select toxins, for each strain listed. Please refer to the instructions and guidance documents for Form 2. If approved, note that an amendment to the recipient entity’s registration for the receiving PI’s work objective must be submitted to FSAP.

Experiment 1:
Yes, the experiment involves the deliberate formation of synthetic or recombinant DNA containing genes for the biosynthesis of a select toxin if the select toxin is lethal for vertebrates at an LD[50]<100ng/kg body weight. A formal request to perform this work should be submitted to the Federal Select Agent Program.

Experiment 2:
No, this is not considered a restricted experiment, but the construct (product of the experiment) is subject to the select agent regulations. Although wild-type E.coli strains alone are not considered select agents and the expected yield of toxin is less than the regulated amount, since the recombinant E. coli strain contains the nucleic acids that encode for and express a toxic form of the toxin lethal for vertebrates at an LD[50] < 100 ng/kg body weight, the recombinant strain is now considered a select agent and subject to select agent regulations. This work must be included in the registration application.