Institutional Biosafety Committee (IBC) review/approval is required prior to the initiation of both exempt and non-exempt experiments.

The information below is a brief description of NON-EXEMPT and EXEMPT experiments from the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules. 

NON-EXEMPT EXPERIMENTS- RECOMBINANT OR SYNTHETIC NUCLEIC ACID MOLECULES

• Section III-A: Studies that involve the deliberate transfer of drug resistance to microorganisms (not known to acquire the trait naturally) that can compromise the use of the drug to control the microorganism and its disease in humans, veterinary medicine or agriculture.
  • Requires NIH Director and IBC oversight
  • Example: Transferring a drug resistance trait that is used, had previously been used, may be used (outside the U.S.), or that is related to other drugs that are used to treat or control disease agents. These include: Transfer of Erythromycin resistance into Borrelia burgdorferi; Transfer of Pyrimethamine resistance into Toxoplasma gondii; Transfer of Chloramphenicol resistance into Rickettsia conorii; Transfer of Tetracycline resistance into Porphyromonas gingivalis.
• Section III-B: Studies that involve the deliberate formation of recombinant or synthetic nucleic acid molecules 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. microbial toxins such as the botulinum toxins, tetanus toxin, diphtheria toxin, and Shigella dysenteriae neurotoxin).
  • Requires NIH/OSP and IBC oversight.
  • Example: Cloning toxin genes (or using plasmids that express genes that encode toxins with low LD₅₀s) such as Botulinum, Tetrodotoxin, Ricin, T-2, Saxitoxin, Abrin, Tetanus, Shigella dysenteriae, Pertussis, Staph Aureus Beta, Shiga Toxin, and Conotoxins.
• Section III-C: Gene transfer experiments in humans.
  • Requires IRB and IBC oversight
  • Example: Use of a defective adenoviral vector to deliver the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene intranasally into patients with Cystic Fibrosis; Introduction of a Herpes Simplex Virus- Thymidine Kinase (HSV-TK) transduced cell line into patients with epithelial ovarian carcinoma, followed by therapy with Ganciclovir.
• Section III-D: Requires IBC oversight
  • D-1: Experiments involving the introduction of recombinant or synthetic nucleic acid molecules into Risk Group 2 organisms (including adenovirus and murine retroviral vectors) or higher.
    • Example: Using adenovirus, adenovirus-luciferase or adeno-associated virus to transfect cells. Typically involves use of pathogens or defective pathogen vectors (with or without helper virus), such as adenovirus, adeno-associated virus, Baculovirus, Herpes virus, lentivirus, retrovirus, vaccinia and vesicular stomatitis virus, shigella, salmonella, yersinia, and E. histolytica.
  • D-2: Experiments in which DNA from human or animal pathogens (Risk Group 2 and higher) is introduced into a non-pathogenic host-vector system.
    • Example: Yersinia pseudotuberculosis genes encoding outer membrane adhesions are cloned into plasmid vectors for re-introduction into mutant strains of the same bacteria or E. coli.
  • D-3: Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses (Risk Group 2 or higher) in the presence of helper virus in tissue culture systems.
    • Example: Insertion of Kaposi’s Sarcoma-associated Herpesvirus (KSHV) genes into defective lentiviral vectors.
  • D-4: Experiments involving whole animals in which the animal's genome has been altered by stable introduction of recombinant or synthetic nucleic acid molecules, or nucleic acids derived therefrom, into the germ-line and experiments involving viable recombinant or synthetic nucleic acid molecule-modified microorganisms tested on whole animals. (This section does not include generation or breeding transgenic rodents, see Section III-E). For more information, see Animal experiments covered under the NIH guidelines.
    • Requires IACUC and IBC oversight.
    • Example: Creation of transgenic animals (mice, rats, zebrafish, drosophila, etc.) or knockout animals that leave genetic material in the animal as part of the silencing of the gene. 
    • Note: the purchase (or transfer to your lab) of previously created transgenic rodents is exempt from the regulations.
  • D-5 (& E2): Experiments to genetically engineer plants by recombinant or synthetic nucleic acid molecule methods, to use such plants for other experimental purposes (e.g., response to stress), to propagate such plants, or to use plants together with microorganisms or insects containing recombinant or synthetic nucleic acid molecules (typically risk group 2 or higher: BSL2 or BSL2-P).
    • Example: Creation of transgenic plants; Inserting a gene for herbicide tolerance in food or ornamental plants.
  • D-6: Experiments involving more than 10 Liters of culture of organisms containing recombinant or synthetic nucleic acids molecules.
    • Example: Use of a 10-liter fermentor or growing up to five 2-liter flasks of recombinant or synthetic nucleic acid molecule culture (i.e. E. coli K-12) qualifies as a large scale experiment.
  • D-7: Experiments involving influenza viruses generated by recombinant or synthetic methods.
  • D-8: Experiments involving Gene Drive Modified Organisms
    • Example: Using gene drives to genetically modify mosquitos to include a new trait that would no longer allow it to transmit the pathogen that causes malaria.
    • This type of research must be conducted at a minimum of Biosafety Level 2.
• Section III-E: Requires IBC oversight
  • E-1: Experiments involving the formation of recombinant or synthetic nucleic acid molecules containing no more than two-thirds of the genome of any eukaryotic virus (cells must lack helper virus for the families of defective viruses used). This category is used for all routine recombinant or synthetic nucleic acid cloning or gene expression with low risk agents (e.g., E. coli cloning strains).
    • Example: Inserting DNA sequences that encode reporters that are measured (lacZ, luciferase, eGFP, dsRed2, etc.), or that encode enzymes that are potentially therapeutic (nitric oxide synthases, superoxide dismutase, siRNA) against mRNAs that promote disease, etc. into viral vectors that retain no more than 2/3 of the original viral genomic sequence. The cDNAs will be driven by the following promoter: CMV IE, RSV LTR, and cardiac Troponin T (cTnT).
  • E-2 (& D-4): Experiments involving nucleic acid molecule-modified whole plants, and/or experiments involving recombinant or synthetic nucleic acid molecule-modified organisms associated with whole plants (typically group 1: BSL1 or BSL1-P).
  • E-3: Experiments involving the generation of risk group 1 (e.g., ABSL-1) rodents in which the animal's genome has been altered by stable introduction of recombinant or synthetic nucleic acid molecules, or nucleic acids derived therefrom, into the germ-line (transgenic rodents). See III-D4 for experiments requiring BSL-2 or higher containment and practices.
    • Requires IACUC and IBC oversight.

EXEMPT (Section III-F) EXPERIMENTS- RECOMBINANT OR SYNTHETIC NUCLEIC ACID MOLECULES

Experiments that:

• F1: Include synthetic nucleic acids that:
  • Can neither replicate nor generate nucleic acids that can replicate in any living cell; and
  • Are not designed to integrate into DNA; and
  • Do not produce a toxin that is lethal for vertebrates at an LD₅₀ of less than 100 nanograms per kilogram body weight?
• F-2: Experiments that are:
  • ​Not in organisms, cells, or viruses; and
  • That have not been modified or manipulated to render them capable of penetrating cellular membranes?
• F-3: Consist solely of the exact recombinant or synthetic nucleic acid sequences from a single source that exists contemporaneously in nature?
• F-4: Consist entirely of nucleic acids from a prokaryotic host including its indigenous plasmids or vectors when propagated only in that host (or a closely related strain of the same species), or when transferred to another host by well-established physiological means?
• F-5: Consist entirely of nucleic acids from an eukaryotic host including chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or a closely related strain of the same species)?
• F-6: Consist entirely of DNA segments from different species that exchange DNA by known physiological processes, though one or more of the segments may be a synthetic equivalent?
• F-7: With those genomic DNA molecules that have acquired a transposable element, provided the transposable element does not contain any recombinant and/or synthetic DNA?
• F-8: Do not present a significant risk to health or the environment as determined by the NIH Director, including:
  • ​C-I: Using recombinant or synthetic nucleic acid molecules containing less than one-half of any eukaryotic viral genome that are propagated and maintained in cells in tissue culture?
  • C-II: Using Escherichia coli K-12 host-vector systems?
  • C-III: Using Saccharomyces cerevisiae and Saccharomyces uvarum host-vector systems?
  • C-IV: Using Kluyveromyces lactis host-vector systems?
  • C-V: Using Bacillus subtilis or Bacillus licheniformis host-vector systems?
  • C-VI: Using extrachromosomal elements of gram positive organisms?
  • C-VII: Purchasing or transferring transgenic rodents (BSL-1 only)?
  • C-VIII: Generating BSL-1 transgenic rodents via breeding? 

Additional Information:

NIH/OSP FAQs

Biosafety Considerations for research with lentiviral vectors

Animal experiments covered under the NIH Guidelines