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NIH Guidelines Reference & Summary Guide

Compliance Policy Overview

Compliance with the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules is mandatory for all institutions receiving NIH funding for recombinant DNA (rDNA) research. Principal Investigators (PIs) are ultimately responsible for ensuring their laboratories remain fully compliant.

Official NIH reference links:

Below is a summary of experiment categories covered under the NIH Guidelines to help identify which requirements apply to your research. Note: Please consult the official NIH links above for the most up-to-date regulations.

Major Categories of Experiments under NIH Guidelines

The NIH Guidelines categorize research with recombinant and synthetic nucleic acid molecules into major
sections based on potential risk and required oversight levels. Below is the structured breakdown of these
regulatory categories:

RAC - Recombinant DNA Advisory Committee

IRB - Institutional Review Board

NIH OSP - Office of Science Policy, former Office of Biotechnology Activities (OBA)

RG - Risk Group

BSL- Bio Safety Level

ABSL - Animal Biosafety Level

Section Subsection Approval Requirements
 Description & Scope
 Examples/ Comments

Section  III-A

(not allowed at MS&T) Requires IBC Approval, RAC
Review, and NIH Director
Approval Before Initiation		 Deliberate transfer of drug
resistance traits to microorganisms not known to
acquire them naturally, where
such acquisition could
compromise disease control in
human/veterinary medicine or
agriculture.		 Cloning rifampin resistance
into Mycobacterium
tuberculosis; cloning
tetracycline resistance into
Chlamydia trachomatis.

Section  III-B

 (not allowed at MS&T) Requires IBC and NIH OSP
(OBA) Approval Before
Initiation		 Experiments involving
deliberate cloning of toxin
molecules with an LD50 of less
than 100 ng/kg body weight.		 Work involving Botulinum
toxins, Tetanus toxin,
Diphtheria toxin, or Shigella
dysenteriae neurotoxin.

Section  III-C

 (not allowed at MS&T) Requires IBC Approval, IRB
Approval, and RAC Review
Before Initiation		 Deliberate transfer of rDNA or
synthetic nucleic acids (or
derived DNA/RNA) into one or
more human research
participants (Human Gene
Therapy).		 Clinical trials utilizing viral
vectors or modified cellular
therapies in human subjects
(Appendix M requirements
apply).
Section III-D, III-E, III-F The majority of the rDNA at MS&T falls into one of those sections. The information regarding the various applications in these categories is present in the table format on the following pages for quick reference.
Section III-D

 

Requires IBC Approval Prior to

initiation

 Experiments using Risk Group
2, 3, 4, or Restricted Agents;
infectious/defective DNA/RNA
virus host-vector systems;
transgenic animals or plants.		 Cloning RG2/RG3 genes into
prokaryotic systems, viral
vector production (lentivirus,
adenovirus), transgenic mouse
work.
III-D-1

 

 

 Experiments using Risk Group (RG) 2, 3, or 4, or Restricted Agents as host-vector systems USDA/APHIS permit may be required for work with certain plant or animal pathogens. See USDA/APHIS
 

III-D-1-a

rDNA into RG2 agents Usually conducted at BSL-2/ABSL-2
 

III-D-1-b

(not allowed at MS&T)

rDNA into RG3 agents Usually conducted at BSL-3/ABSL-3
 

III-D-1-c

(not allowed at MS&T)

rDNA into RG4 agents Usually conducted at BSL-4/ABSL-4
 

III-D-1-d

(not allowed at MS&T)

rDNA into restricted agents Containment determined on a case-by-case basis following NIH/OBA review

III-D-2

 

Experiments in which DNA from Risk Group 2, 3, or 4, or Restricted Agents is cloned into nonpathogenic prokaryotic or lower eukaryotic host-vector systems  
 

III-D-2-a

rDNA from RG2, RG3, or RG4 agents

BSL-2 containment for cloning from RG2 or RG3 pathogens; specific lowering to BSL-1 may be approved by the IBC

BSL-2 for cloning from RG4 agents only after demonstration that a totally & irreversibly defective fragment of the agent’s genome is present in a given recombinant; otherwise BSL-4 required.

 

III-D-2-b

rDNA from restricted agents Containment determined on a case-by-case basis following NIH/OBA review

III-D-3

Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses in the presence of helper virus in tissue culture systems  

 

III-D-3-a

Infectious/defective RG2 viruses with helper virus Usually conducted at BSL-2

 

III-D-3-b

(not allowed at MS&T)

Infectious/defective RG3 viruses with helper virus Usually conducted at BSL-3

 

III-D-3-c

(not allowed at MS&T)

Infectious/defective RG4 viruses with helper virus Usually conducted at BSL-4

 

III-D-3-d

(not allowed at MS&T)

Infectious/defective restricted poxviruses with helper virus Containment determined on a case-by-case basis following NIH/OBA review

 

III-D-3-e

Viruses not covered in III-D-3-a through III-D-3-d Usually conducted at BSL-1

III-D-4

 

Experiments involving whole animals in which the genome has been altered by stable introduction into the germ-line (transgenic animals) and experiments involving viable rDNA-modified microorganisms tested on whole animals  

 

III-D-4-a

rDNA from any source except for > 2/3 of eukaryotic viral genome Usually conducted at BSL-1; viral vectors must not lead to transmissible infection

 

III-D-4-b

rDNA involving whole animals and not covered by III-D-1 or III-D-4-a Appropriate containment decided by the IBC

 

III-D-4-c (1)

Generating transgenic rodents that require BSL-1 containment Covered under Section III-E-3

 

III-D-4-c (2)

Purchase or transfer of transgenic rodents Exempt under Section III-F-6, Appendix C-VI

III-D-5

  

Experiments involving whole plants – genetically engineering plants by rDNA methods, using or propagating such plants, using plants with microorganisms or insects containing rDNA  

 

III-D-5-a

(not allowed at MS&T)

Exotic plant pathogens with recognized potential for serious detrimental impact on ecosystems Usually conducted at BSL-2+P/BSL-3P

 

III-D-5-b

(not allowed at MS&T)

 

Readily transmissible exotic agents in which the complete and functional genome may be reconstituted in planta Usually conducted at BSL-2+P/BSL-3P

 

III-D-5-c

(not allowed at MS&T)

 

Readily transmissible exotic agents in the presence of their arthropod vector Usually conducted at BSL-4P

 

III-D-5-d

(not allowed at MS&T)

 

Sequences encoding potent vertebrate toxins introduced into plants or associated organisms Usually conducted at BSL-3P

 

III-D-5-e

(not allowed at MS&T)

 

Microbial pathogens of insects or small animals associated with plants Usually conducted at BSL-2+P/BSL-3P

III-D-6

 

Experiments involving > 10 liters of culture Containment to be decided by IBC; see Appendix K for containment conditions

III-D-7

 

Experiments involving influenza viruses Conducted at the containment level corresponding to the RG of the virus that is the source of the majority of segments

 

III-D-7-a

(not allowed at MS&T)

 

Human H2N2 (1957-1968) BSL-3+ for viruses containing H2 hemagglutinin (HA) segment

BSL-2 for H2 HA gene in cold-adapted, live attenuated vaccine strains and for H2N2 genes other than HA

 

III-D-7-b

(not allowed at MS&T)

Highly pathogenic avian influenza H5N1 Usually conducted at BSL-3+

 

III-D-7-c

(not allowed at MS&T)

 

1918 H1N1 Usually conducted at BSL-3+

 

III-D-7-d

(not allowed at MS&T)

 

Antiviral susceptibility for genes from viruses in III-D-7-a through III-D-7-c Higher containment may be required if any of the genes are resistant to both classes of current antiviral agents (adamantanes and neuraminidase inhibitors)
Section III-E

 

Requires IBC Notice at

Initiation

 Experiments involving low-risk
nucleic acid construction,
limited host-vector systems, or
specific non-pathogenic
organisms.		 Certain low-risk viral vector or
plasmid constructions not
covered under Section III-D or
exempt under III-F.
III-E-1

 

 

 Formation of rDNA molecules containing no more than 2/3 of the genome of any eukaryotic virus BSL-1, provided that cells lack helper virus for the specific families of defective virus being used
III-E-2

 

 

 rDNA-modified plants or rDNA-modified microorganisms associated with plants not covered in sections III-A, III-B, III-D or III-F Usually BSL-1+P/BSL-2P
III-E-3

 

 

 Creation of transgenic rodents  BSL-1; experiments requiring BSL-2 or higher covered under section III-D-4
 

III-E-3-a

 

 Breeding of BSL-1 transgenic rodents Exempt under Section III-F-6, Appendix C-VII
Section III-F   Exempt Experiments Synthetic nucleic acids that
cannot replicate, do not
integrate, and do not encode
lethal toxins (LD50 < 100
ng/kg); standard PCR; specific
non-pathogenic host-vector
systems.		 Use of commercial
oligonucleotides, non-viral
synthetic DNA not integrating
into genomes.
  App C-1   rDNA containing < ½ of any eukaryotic viral genome in tissue culture Exceptions: Experiments described in Sections III-A or III-B, those involving RG3, 4, or restricted agents, large-scale experiments, or cloning of toxin molecule genes coding for biosynthesis of molecules toxic for vertebrates
  App C-II   E. coli K-12 host-vector systems Same exceptions as C-1
  App C-III   Saccharomyces host-vector systems Same exceptions as C-1
  App C-IV   Bacillus subtilis or Bacillus licheniformis host-vector systems Same exceptions as C-1
  App C-V   Extrachromosomal elements of listed gram-positive organisms propagated and maintained in gram-positive organisms See App C-V for list. Same exceptions as C-1
  App C-VI   Purchase or transfer of transgenic rodents Only applies to rodents requiring BSL-1 containment
  App C-VII   Breeding of two different transgenic rodents or breeding of a transgenic rodent with a non-transgenic rodent Requirements:

1) Both parental rodents require BSL-1 containment,

2) Neither parental rodent contains incorporation of > ½ of the genome of an exogenous eukaryotic virus or incorporation of a transgene under control of a gammaretroviral long terminal repeat (LTR) and,

3) Rodent resulting from the breeding not expected to contain > ½ of an exogenous viral genome