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Federal Register / Vol. 46, No. 126 / Wednesday. July 1, 1981 / Notices 
DEPARTMENT OF HEALTH AND 
HUMAN SERVICES 
National Institutes of Health 
Guidelines for Research Involving 
Recombinant DNA Molecules June 
1981 
These NIH guidelines supersede those 
of November 1980, and will be in effect 
until further notice. 
Table of Contents 
I. Scope of the Guidelines 
I-A — Purpose 
I-B — Definition of Recombinant DNA 
Molecules 
I-C — General Applicability (see IV-B) 
1-D — Prohibitions 
I-E — Exemptions 
I- F — General Definitions (see IV-C) 
H. Containment 
II- A — Standard Practices and Training 
II-B — Physical Containment Levels 
U-B-1 — PI Level 
D-B^l-a — Laboratory Practices 
D-B-l-b — Containment Equipment 
D-B-l-c — Special Laboratory Design 
n-B-2— P2 Level 
II-B-2-a — Laboratory Practices 
U-B-2-b— Containment Equipment 
n-B-2-c — Special Laboratory Design 
D-B-3 — P3 Level 
n-B-3-a — Laboratory Practices 
II-B-3-b — Containment Equipment 
n-B-3-c — Special Laboratory Design 
II-B-4 — P4 Level 
II-B-4-a — Laboratory Practices 
U-B-4-b — Containment Equipment 
n-B-4-c — Special Laboratory Design 
U-C — Shipment 
D-D — Biological Containment 
D-D-l — Levels of Biological Containment 
n-D-l-a— HVl 
n-B-l-b— HV2 
Il-Ef-l-c— HV3 
II- D-2 — Certification of Host-Vector 
Systems 
D-D-2-a — Responsibility 
U-D-2-b— Data To Be Submitted for 
Certification 
n-D-3 — Distribution of Certified Host- 
Vectors 
III. Containment Guidelines for Covered 
Experiments 
III- Ot — Classification of Experiments Using 
Certain Host-Vector Systems 
III-O-l — Experiments Involving Class 3 
Organisms 
III-0-2 — Experiments Involving 
Prokaryotes Nonpathogenic for Man, 
Animals or Plants, and/or Lower 
Eukaryotes Nonpathogenic for Man, 
Animals or Plants 
lU-A — Classification of Experiments Using 
Certain HVl and HV2 Host-Vector 
Systems 
IlI-A-1 — Shotgun Experiments 
IIl-A-l-a — Eukaryotic DNA Recombinants 
III-A-l-b — Prokaryotic DNA 
Recombinants 
UI-A-2-a — Viruses of Eukaryotes 
III-A-2-b — Eukaryotic Organelle DNAs 
UI-A-2-C — Prokaryotic Plasmid and Phage 
DNAs 
IIl-A-3 — Lowering of Containment Levels 
for Characterized or Purified DNA 
Preparations and Clones 
III-A-3-a — Purified DNA Other than 
Plasmids, Bacteriophages, and Other 
Viruses 
Ul-A-3-b — Characterized Clones of DNA 
Recombinants 
III-B — Experiments with Prokaryotic Host- 
Vectors Other than E. coli K-12 
III-B-1 — HVl and HV2 Systems 
III-B-2 — Return of DNA Segments to 
Prokaryotic Non-HVl Host or Origin 
III-B-3 — Non-HVl Systems 
III-C — Experiments with Eukaryotic Host- 
Vectors 
IlI-C-1 — Vertebrate Host-Vector Systems 
Ill-C-l-a — Polyoma Virus 
Ill-C-l-b — Simian Virus 40 
IIl-C-l-c — Human Adenoviruses 2 and 5 
ni-C-l-d — Murine Adenovirus Strain FL 
IlI_C_l_e— All Viral Vectors 
Ill-C-l-f — Nonviral Vectors 
III-C-2 — Invertebrate Host-Vector Systems 
III-C-2-a — Insect Viral Vectors 
III-C-2-b — Nonviral Vectors 
lII-C-3 — Plant Viral Host-Vector Systems 
III-C-4 — Plant Host-Vector Systems Other 
than Viruses 
III-C-5 — Fungal or Similar Lower 
Eukaryotic Host-Vector Systems 
III-C-6 — Return of DNA Segments to a 
Higher Eukaryotic Host of Origin 
ni-C-7 — Transfer of Cloned DNA 
Segments to Eukaryotic Organisms 
III-C-7-a — ^Transfer to Non-human 
Vertebrates 
III-C-7-b— Transfer to Higher Plants 
III- D — Complementary DNAs 
UI-E — Synthetic DNAs 
IV. Roles and Responsibilities 
rV-A — Policy 
IV- B — General Applicability 
rV-C — General Definitions 
rV-D — ^Responsibilities of the Institution 
IV-D-1— ‘(General) 
IV-D-2 — ^Membership and Procedures of 
thelBC 
IV-D-3 — Functions of the IBC 
IV-D-4 — Biological Safety Officer 
IV-D-5 — Principal Investigator 
rV-D-5-a — PI— General 
IV-D-5-b — Submissions by the PI to NIH 
IV-D-5-C — Submissions by the PI to the 
IBC 
rV-D-5-d — PI Responsibilities After 
Approval but Prior to Initiating the 
Research 
IV-D-5-e — PI Responsibilities During the 
Conduct of the Approved Research 
IV-E — Responsibilities of NIH 
IV-E-1 — Director 
IV-E-l-a — General Responsibilities of the 
Director, NIH 
IV-E-l-b — Specific Responsibilities of the 
Director, NIH 
IV-E-2 — Recombinant Advisory 
Committee 
IV-E-3 — The Office of Recombinant DNA 
Activities 
IV-E-4 — Other NIH Components 
IV-G — Compliance 
V. Footnotes and References 
VI. - Voluntary Compliance 
VI-A — Basic Policy 
VI-B — IBC Approval 
VI-D — Certification of Host-Vector 
Systems 
Vl-E — Requests for Exceptions, 
Exemptions, Approvals 
Vl-F — Protection of Proprietary Data 
Appendix A — Exemptions Under I-E-4 
Appendix B — Classification of 
Microorganisms on the Basis of Hazard 
Appendix C — Exemptions Under I-E-5 
Appendix D — HVl and HV2 Host-Vector 
Systems Assigned Containment Levels as 
Specified in the Subsections of Section 111- 
A 
Appendix E — Actions Taken Under the 
Guidelines 
Appendix F — Certified Host-Vector Systems 
Appendix G — Containment Conditions for 
Cloning of Genes Coding for the 
Biosynthesis of Toxins for Vertebrates 
Appendix H — Experiments Covered by 
Section Ill-O 
I. Scope of the Guidelines 
I-A. Purpose. The purpose of these 
Guidelines is to specify practices for 
construction and handling (i) 
recombinant DNA molecules and (ii) 
organisms and viruses containing 
recombinant DNA molecules. 
I-B. Definition of Recombinant DNA 
Molecules. In the context of these 
Guidelines, recombinant DNA molecules 
are dehned as either (i) molecules which 
are constructed outside living cells by 
joining natural or synthetic DNA 
segments to DNA molecules that can 
replicate in a living cell, or (ii) DNA 
molecules that result from the 
replication of those described in (i) 
above. 
I-C. General Applicability. See 
Section IV-B. 
I-D. Prohibitions. The following 
experiments are not to be initiated at the 
present time: 
I-D-1. Formation of recombinant 
DNAs derived from the pathogenic 
organisms classified (7) as Class 4 or 5 
(2) or from cells known [2 A) to be 
infected with such agents, regardless of 
the host-vector system used. 
I-D-2. Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxins lethal for 
vertebrates at an LDm of less than 100 
nanograms per kilogram body weight 
(e.g., the botulinum toxins, tetanus toxin, 
diphtheria toxin. Shigella dysenteriae 
neurotoxin). Guidelines for the cloning 
of DNAs containing genes coding for the 
biosynthesis of toxins which are lethal 
to verebrates at 100 nanograms to 100 
micrograms per kilogram body weight 
are specified in Appendix G, which 
overrides other parts of the Guidelines 
(e.g., exemptions, return to host of 
origin, etc.). 
I-D-3. (Deleted] 
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