APPENDIX C. EXEMPTIONS
UNDER SECTION III-F-6
Appendix C-I. Recombinant DNA in
Tissue Culture
Appendix C-II. Escherichia coli K-12 Host-Vector Systems
Appendix C-III. Saccharomyces Host-Vector Systems
Appendix C-IV. Kluyveromyces Host-Vector Systems
Appendix C-V. Bacillus subtilis or Bacillus
licheniformis Host-Vector Systems
Appendix C-VI. Extrachromosomal Elements
of Gram Positive Organisms
Appendix C-VII. The Purchase or Transfer of
Transgenic Rodents
Appendix C-VIII. Generation of BL1 Transgenic Rodents via Breeding.
Appendix C-IX. Footnotes and References of
Appendix C
Section III-F-6 states that exempt from these NIH Guidelines
are "those that do not present a significant risk to health or the
environment (see Section
IV-C-1-b-(1)-(c), NIH Director--Specific Responsibilities), as
determined by the NIH Director, with the advice of the RAC, and following
appropriate notice and opportunity for public comment. See Appendix C, Exemptions under Sections
III-F-6, for other classes of experiments which are exempt from the NIH
Guidelines." The following
classes of experiments are exempt under Section
III-F-6:
Recombinant DNA molecules containing less than one-half of
any eukaryotic viral genome (all viruses from a single family being considered
identical -- see Appendix C-IX-E, Footnotes
and References of Appendix C), that are propagated and maintained in cells
in tissue culture are exempt from these NIH Guidelines with the
exceptions listed in Appendix C-I-A.
The following categories are not exempt from the NIH
Guidelines: (i) experiments described in Section
III-B which require NIH/OBA and Institutional Biosafety Committee approval
before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or
restricted organisms (see Appendix B, Classification
of Human Etiologic Agents on the Basis of Hazard, and Sections
V-G and V-L, Footnotes and References of Sections I through IV) or
cells known to be infected with these agents, (iii) experiments involving the
deliberate introduction of genes coding for the biosynthesis of molecules that
are toxic for vertebrates (see Appendix
F, Containment Conditions for Cloning of Genes Coding for the
Biosynthesis of Molecules Toxic for Vertebrates), and (iv) whole plants
regenerated from plant cells and tissue cultures are covered by the exemption
provided they remain axenic cultures even though they differentiate into
embryonic tissue and regenerate into plantlets.
Experiments which use Escherichia coli K-12
host-vector systems, with the exception of those experiments listed in Appendix
C-II-A, are exempt from the NIH Guidelines provided that: (i) the Escherichia coli host
does not contain conjugation proficient plasmids or generalized transducing
phages; or (ii) lambda or lambdoid or Ff bacteriophages or non-conjugative
plasmids (see Appendix C-IX. Footnotes and References of Appendix C, Footnotes and References
of Appendix C) shall be used as vectors.
However, experiments involving the insertion into Escherichia coli
K-12 of DNA from prokaryotes that exchange genetic information (see Appendix C-IX. Footnotes and References of Appendix C, Footnotes and References of Appendix C)
with Escherichia coli may be performed with any Escherichia
coli K-12 vector (e.g., conjugative plasmid). When a non-conjugative vector is used, the Escherichia coli
K-12 host may contain conjugation-proficient plasmids either autonomous or
integrated, or generalized transducing phages.
For these exempt laboratory experiments, Biosafety Level (BL) 1 physical
containment conditions are recommended.
For large-scale fermentation experiments, the appropriate physical
containment conditions need be no greater than those for the host organism
unmodified by recombinant DNA techniques; the Institutional Biosafety Committee
can specify higher containment if deemed necessary.
The following categories are not exempt from the NIH
Guidelines:
(i) experiments described in Section
III-B which require NIH/OBA and Institutional Biosafety Committee approval
before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or
restricted organisms (see Appendix B,
Classification of Human Etiologic Agents on the Basis of Hazard, and Sections
V-G and V-L, Footnotes and References of Sections I through IV) or
cells known to be infected with these agents may be conducted under containment
conditions specified in Section
III-D-2 with prior Institutional Biosafety Committee review and approval,
(iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv)
experiments involving the cloning of toxin molecule genes coding for the
biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes
Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
Experiments involving Saccharomyces cerevisiae and Saccharomyces
uvarum host-vector systems, with the exception of experiments listed in
Appendix C-III-A, are exempt from the NIH Guidelines. For these exempt experiments, BL1 physical
containment is recommended. For
large-scale fermentation experiments, the appropriate physical containment
conditions need be no greater than those for the host organism unmodified by
recombinant DNA techniques; the Institutional Biosafety Committee can specify
higher containment if deemed necessary.
The following categories are not exempt from the NIH
Guidelines:
(i) experiments described in Section
III-B which require NIH/OBA and Institutional Biosafety Committee approval
before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or
restricted organisms (see Appendix B,
Classification of Human Etiologic Agents on the Basis of Hazard, and Sections
V-G and V-L, Footnotes and References of Sections I through IV) or
cells known to be infected with these agents may be conducted under containment
conditions specified in Section
III-D-2 with prior Institutional Biosafety Committee review and approval,
(iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv)
experiments involving the deliberate cloning of genes coding for the
biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes
Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
Experiments involving Kluyveromyces lactis, host-vector systems, with the exception of experiments listed in Appendix C-IV-A, are exempt from the NIH Guidelines provided laboratory-adapted strains are used (i.e. strains that have been adapted to growth under optimal or defined laboratory conditions). For these exempt experiments, BL1 physical containment is recommended. For large-scale fermentation experiments, the appropriate physical containment conditions need be no greater than those for the host organism unmodified by recombinant DNA techniques; the Institutional Biosafety Committee may specify higher containment if deemed necessary.
The following categories are not exempt from the NIH
Guidelines:
(i) experiments described in Section
III-B which require NIH/OBA and Institutional Biosafety Committee approval
before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or
restricted organisms (see Appendix B,
Classification of Human Etiologic Agents on the Basis of Hazard, and Sections
V-G and V-L, Footnotes and References of Sections I through IV) or
cells known to be infected with these agents may be conducted under containment
conditions specified in Section
III-D-2 with prior Institutional Biosafety Committee review and approval,
(iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv)
experiments involving the deliberate cloning of genes coding for the
biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes
Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
Any asporogenic Bacillus subtilis or asporogenic
Bacillus licheniformis strain which does not revert to a spore-former with
a frequency greater than 10-7 may be used for cloning DNA with the
exception of those experiments listed in Appendix C-V-A, Exceptions. For these exempt laboratory experiments, BL1
physical containment conditions are recommended. For large-scale fermentation experiments, the appropriate
physical containment conditions need be no greater than those for the host
organism unmodified by recombinant DNA techniques; the Institutional Biosafety
Committee can specify higher containment if it deems necessary.
The following categories are not exempt from the NIH Guidelines: (i) experiments described in Section III-B which require NIH/OBA and Institutional Biosafety Committee approval before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or restricted organisms (see Appendix B, Classification of Human Etiologic Agents on the Basis of Hazard, and Sections V-G and V-L, Footnotes and References of Sections I through IV) or cells known to be infected with these agents may be conducted under containment conditions specified in Section III-D-2 with prior Institutional Biosafety Committee review and approval, (iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv) experiments involving the deliberate cloning of genes coding for the biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
Recombinant DNA molecules derived entirely from
extrachromosomal elements of the organisms listed below (including shuttle
vectors constructed from vectors described in Appendix
C), propagated and maintained in organisms listed below are exempt from
these NIH Guidelines.
Bacillus amyloliquefaciens
Bacillus amylosacchariticus
Bacillus anthracis
Bacillus aterrimus
Bacillus brevis
Bacillus cereus
Bacillus globigii
Bacillus licheniformis
Bacillus megaterium
Bacillus natto
Bacillus niger
Bacillus pumilus
Bacillus sphaericus
Bacillus stearothermophilis
Bacillus subtilis
Bacillus thuringiensis
Clostridium acetobutylicum
Lactobacillus casei
Listeria grayi
Listeria monocytogenes
Listeria murrayi
Pediococcus acidilactici
Pediococcus damnosus
Pediococcus pentosaceus
Staphylococcus aureus
Staphylcoccus carnosus
Staphylococcus epidermidis
Streptococcus agalactiae
Streptococcus anginosus
Streptococcus avium
Streptococcus cremoris
Streptococcus dorans
Streptococcus equisimilis
Streptococcus faecalis
Streptococcus ferus
Streptococcus lactis
Streptococcus ferns
Streptococcus mitior
Streptococcus mutans
Streptococcus pneumoniae
Streptococcus pyogenes
Streptococcus salivarious
Streptococcus sanguis
Streptococcus sobrinus
Streptococcus thermophylus
The following categories are not exempt from the NIH
Guidelines: (i) experiments described in Section
III-B which require NIH/OBA and Institutional Biosafety Committee approval
before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or
restricted organisms (see Appendix B,
Classification of Human Etiologic Agents on the Basis of Hazard, and Sections
V-G and V-L, Footnotes and References of Sections I through IV) or
cells known to be infected with these agents may be conducted under containment
conditions specified in Section
III-D-2 with prior Institutional Biosafety Committee review and approval,
(iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv)
experiments involving the deliberate cloning of genes coding for the
biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes
Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
The purchase or transfer of transgenic rodents for
experiments that require BL1 containment (See Appendix
G-III-M, Footnotes and References of Appendix G) are exempt from the
NIH Guidelines.
The breeding of two different transgenic rodents or the breeding of a transgenic rodent and a non-transgenic rodent with the intent of creating a new strain of transgenic rodent that can be housed at BL1 containment will be exempt from the NIH Guidelines if:
(1) Both parental rodents can be housed under BL1 containment; and
(2) neither parental transgenic rodent contains the following genetic modifications: (i) incorporation of more than one-half of the genome of an exogenous eukaryotic virus from a single family of viruses; or (ii) incorporation of a transgene that is under the control of a gammaretroviral long terminal repeat (LTR); and
(3) the transgenic rodent that results from this breeding is not expected to contain more than one-half of an exogenous viral genome from a single family of viruses.
Appendix C-IX-A. The NIH Director, with advice of the RAC,
may revise the classification for the purposes of these NIH
Guidelines (see Section
IV-C-1-b-(2)-(b), Minor Actions).
The revised list of organisms in each Risk Group is located in Appendix B.
Appendix C-IX-B.
A subset of non-conjugative plasmid vectors are poorly mobilizable
(e.g., pBR322, pBR313). Where
practical, these vectors should be employed.
Appendix C-IX-C.
Defined as observable under optimal laboratory conditions by
transformation, transduction, phage infection, and/or conjugation with transfer
of phage, plasmid, and/or chromosomal genetic information. Note that this definition of exchange may be
less stringent than that applied to exempt organisms under Section
III-F-5, Exempt Experiments.
Appendix C-IX-D.
As classified in the Third Report of the International Committee on
Taxonomy of Viruses: Classification and
Nomenclature of Viruses, R. E. F. Matthews (ed.), Intervirology 12
(129-296), 1979.
Appendix C-IX-E.
i.e., the total of all genomes within a Family shall not exceed one-half
of the genome.