Bio-ARROW help
Antibiotics, herbicides, etc. that are used for selection purposes, even if they are not used to treat infections, should be included in this section.
Examples of how to enter genes fragments used to knock-down or silence genes.
sgRNA used
for CRISPR modifications:
Gene name |
sgRNAs |
Gene source |
Synthetic |
Nature of
insert or protein |
Describe the
function of the sgRNA you are using for CRISPR modifications |
Gene use |
Select
all that apply (e.g., Knockout, Silencing, Up-regulations, etc.) |
Gene use –
other details |
|
Cas9 gene used for CRISPR modifications:
Gene name |
Cas9 |
Gene source |
Streptococcus
pyogenes |
Nature of
insert or protein |
Nuclease |
Gene use |
Cut
nucleic acid |
Gene use –
other details |
|
DNA carried
on a vector that is introduced to upregulate a gene:
Gene name |
Estrogen receptors:
alpha, beta, GPER |
Gene source |
Human,
Non-human primate |
Nature of
insert or protein |
DNA-binding
transcription factor |
Gene use |
Upregulating |
Gene use –
other details |
|
siRNA:
Gene name |
Fragments of
gene X |
Gene source |
Synthetic |
Nature of insert
or protein |
Describe the
function of the gene you are knocking down or silencing |
Gene use |
Knockout Silencing |
Gene use –
other details |
|
Examples on
how to broadly describe a category of genes:
Gene name |
Genes
involved in energy metabolism (e.g., ldh, ackA, pyk, pta) |
Gene source |
E. coli,
yeast |
Nature of
insert or protein |
Metabolic
genes |
Gene use |
Gene
expression Complement
knockout (restore function in trans) |
Gene use –
other details |
|
Gene name |
Virulence
factors (e.g., SpA, Protein A) |
Gene source |
S. aureus |
Nature of
insert or protein |
Virulence
factors |
Gene use |
Downregulating Knockout Complement
knockout (restore function in trans) |
Gene use –
other details |
Do not
anticipate causing increased pathogenicity |
Example of how to describe the use of a genome library:
Gene name |
cDNA library
|
Gene source |
Aspergillus |
Nature of
insert or protein |
cDNA library |
Gene use |
Gene
expression Complement
knockout (restore function in trans) |
Gene use –
other details |
Not
targeting toxin genes |
Gene expression for protein production:
Gene name |
Hsp |
Gene source |
E. coli,
yeast, Drosophila |
Nature of
insert or protein |
Chaperones |
Gene use |
Point
mutation Gene
expression |
Gene use –
other details |
|
Plasmids that are non-conjugative or non-mobilizable lack the nic/bom elements required for mobility. Plasmids that lack F factors are considered nonconjugative (see http://en.wikipedia.org/wiki/F-plasmid for more information).
Agrobacterium plasmids that are disarmed lack the tumor inducing genes needed for pathogenicity.
This section of the biosafety protocol is for information on the use of gene editing tools such as CRISPR/Cas9, TALENs, or ZFNs. It is also used to enter information on organisms or cells you are working with been created using genome editing tools. Even if these genome editing tools are not used directly for editing purposes, they should be added to this section.
If you are using microbes, animals, or plants that were generated by a collaborator or company using CRISPR-based genome editing tools, this should be clearly stated. Any additional information related to the potential risk of gene drive and how it would be mitigated that is not already included elsewhere in the protocol should be described here. Please clarify if the organism being modified would be at an advantage if inadvertently released into the environment. Describe how risk of release will be mitigated. If you are using CRISPR systems for purposes other than genome editing (e.g., CRISPRi, CRISPRa, CETChseq), the components of the editing system and their intended purpose should be clearly described. Many non-traditional uses of CRISPR are based on mutants that lack nuclease activity. Without this activity, there may be no risk of gene drive. Other risks (e.g., oncogenic potential) may still exist, however, and should be addressed.
This section uses branching logic; only questions relevant to the organism or cells being modified will be displayed.
Example table: Gene drive is
not relevant; only in somatic cells where oncogenic potential exists |
|
Organism or Cells Being
Modified |
Human Cells or Cell
Culture (Somatic) |
Targeted Sequence(s) |
Genes involved in
chromatin remodeling |
Nuclease Type |
CAS9 gene (cutting
intact) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
pQCXIN , pCDNA |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Transfection; Other
|
Delivery Method(s) -
Other |
liposome |
Integration of Genome
Editing Components |
No Value Entered |
Integration of Genome
Editing Components - Other |
No Value Entered |
Gene Drive |
No Value Entered |
Gene Drive Additional
Information |
No Value Entered |
|
|
|
|
Example table: Gene drive is
not relevant; only non-sexually reproducing bacteria. |
|
Organism or Cells Being
Modified |
Microbe |
Targeted Sequence(s) |
Genes involved in
biofilm production |
Nuclease Type |
Zinc finger nuclease
(ZFN) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
PZFN-1 |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Bacterial
transformation |
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
No Value Entered |
Integration of Genome
Editing Components - Other |
No Value Entered |
Gene Drive |
No Value Entered |
Gene Drive Additional
Information |
No Value Entered |
|
|
|
|
Example table: Sexually
reproducing organism, gene drive could not occur because purified nuclease
protein used |
|
Organism or Cells Being
Modified |
Animal |
Targeted Sequence(s) |
Genes involved in
insulin production |
Nuclease Type |
Cas9 protein (no DNA,
protein only) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
No value Entered |
Delivery Vehicle(s) -
Other |
Purified gRNA
|
Delivery Method(s) |
Direct injection
|
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
Purified nuclease
protein, will not integrate |
Integration of Genome
Editing Components - Other |
N/A |
Gene Drive |
No |
Gene Drive Additional
Information |
No Value Entered |
|
|
|
|
Example table:
Sexually reproducing organism, separate constructs designed to prevent
integration and potential for gene drive |
|
Organism or Cells Being
Modified |
Animal |
Targeted Sequence(s) |
Genes involved in
superoxide dismutase |
Nuclease Type |
Cas9 nickase gene
(single stranded cut) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
sgRNA, pSuperOx |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Viral transduction
|
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
Separate constructs,
will not integrate at the target site (i.e., flanking sequences not
homologous to sequences near the target site) |
Integration of Genome
Editing Components - Other |
N/A |
Gene Drive |
No |
Gene Drive Additional
Information |
No Value Entered |
|
|
|
|
Example table:
Sexually reproducing organism, single construct designed to allow integration
for purpose of creating gene drive |
|
Organism or Cells Being
Modified |
Animal |
Targeted Sequence(s) |
Genes involved in
fertility |
Nuclease Type |
CAS9 gene (cutting
intact) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
pCRISPR |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Direct injection
|
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
Single construct,
could integrate at the target site (i.e., flanking sequences are homologous
to sequences near the target site and could have the potential to be used as
template for homology-based DNA repair) |
Integration of Genome
Editing Components - Other |
Integration required
for experimental design. |
Gene Drive |
Yes |
Gene Drive Additional
Information |
The purpose of the
study is to enact gene drive to test approach for changing population
dynamics of wild type mosquito. To prevent accidental release, work will be
performed at ACL2 using double containment housing and traps; see Research
Description for additional details. |
|
|
|
|
Example table:
Sexually reproducing organism, single construct designed to allow integration
and nuclease bred out of subsequent generations to avoid gene drive |
|
Organism or Cells Being
Modified |
Plant |
Targeted Sequence(s) |
Genes encoding human
antibodies |
Nuclease Type |
Transcription activator-like effector
nuclease (TALEN) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
pTALEN 5.0 |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Agrobacterium-mediated
gene transfer |
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
Single construct, could integrate at the
target site (i.e., flanking sequences are homologous to sequences near the
target site and could have the potential to be used as template for
homology-based DNA repair) |
Integration of Genome
Editing Components - Other |
The goal is to integrate the genes
(stability) as goal of experimental design. |
Gene Drive |
No |
Gene Drive Additional
Information |
No Value Entered |
|
|
|
|
Example table:
Sexually reproducing organism, gene drive could not occur because using
catalytically inactive nuclease for purposes other than genome editing |
|
Organism or Cells Being
Modified |
Animal |
Targeted Sequence(s) |
Targeting the promoter
of liver enzymes |
Nuclease Type |
dCas9 gene (cannot
cut) |
Nuclease Type - Other |
No Value Entered |
Delivery Vehicle(s) |
pLENTI 2.0 |
Delivery Vehicle(s) -
Other |
No Value Entered |
Delivery Method(s) |
Viral transduction
|
Delivery Method(s) -
Other |
No Value Entered |
Integration of Genome
Editing Components |
Dead nuclease, no gene
drive potential (e.g., dCas9) |
Integration of Genome
Editing Components - Other |
N/A |
Gene Drive |
No |
Gene Drive Additional
Information |
No Value Entered |
Due to revisions to the NIH Guidelines in 2019, the IBC is no longer required to consider requirements for RAC review and reporting of adverse events. Since 2016 new studies no longer need to submit responses to Appendix M to NIH though if your study has this document, please provide.
If you need assistance in completing the biosafety protocol for a human clinical trial, please contact the Office of Biological Safety.
Still have questions? Call the Office of Biological Safety (OBS) at 608-263-2037. We are happy to help.