Genome Editing and Gene Drive at UCI TALENS, ZNF, CRISPR-Cas9 and Meganucleases There are multiple technologies available for creating permanent genomic modifications both in vitro and in vivo. These include, but are not limited to, TALENS (Transcription Activator-Like effector Nucleases), ZNF (Zinc Finger Nuclease mediated DNA Repair), Meganucleases, and CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats )-Cas9. Among the many applications of these technologies, there is the creation of gene drives that can result in a spread of a desired trait through a population. Regardless of the specific application, an IBC approval is required for research involving any of the above gene editing technologies. The information below has been provided as a guidance on what IBC requires while reviewing your IBC application involving gene editing. Section 14.2, include the following information: Describe the Gene Editing Technology (CRISPR-Cas9, ZNF, TALENS, Meganucleases) Describe the target gene, function of the target gene and the desired effect of the target gene in the system (in-vitro/in vivo) Specify whether the technology is used in prokaryotes (Escherichia coli Top 10 cells), eukaryotes (Mice, Drosophila, Mosquitoes, Plant) or cell lines (human/animal/insect/plant cell lines). *Specify whether the technology will target embryos or germ line cells. How is the gene editing technology being delivered (e.g. Nanoparticles, plasmid, Lentivirus, Adeno-Associate virus etc.?) Will the research involve creation of a gene drive experiment? Include your laboratory’s proposed work with gene drives in vertebrates, invertebrates or plants. *No gene editing of the germ line, human embryo or germ cells for clinical application is currently allowed. Gene editing of human embryos and germ cells for scientific purpose may be allowed, but must be evaluated on a case-by-case basis by the appropriate federal and local scientific review committees. Section 14.3, include the following information: Address the potential effects due to accidental worker exposure. If unknown, state that. Points to consider are: Is the guide sequence specific to animals, humans or could it affect both? What are the effects of Off-target effect of the system, if any? How does route of exposure affect outcome ( inoculation versus ingestion versus inhalation)? Containment requirements: BSL1 for non-viral delivery All waste from gene editing experiment must be treated as biohazardous unless the waste is mixed with chemical or radioactive material. For mixed waste, contact UCI EH&S Hazardous waste 949-824-4578 for more information.   BSL2 for viral delivery, use of human and non-human primate cell lines For all gene drive work in plants, animals and arthropods, please contact IBC for research specific containment requirements. Section 15, Applicable NIH Guidelines: If your work involves using gene drive or gene editing technology in vitro, in human, animal, insect or plants cells, the research will fall under III-E. If viral vector ( RG2 or higher) is used, research will fall under III-D. If the procedures results in toxic or infectious product, there will be additional guidelines that need to be checked. References For more information on the transgene being silenced or over-expressed: Gene CARDS: www.genecards.org/ For more information on potential human outcomes from accidental silencing. JAX Mouse Genome Informatics: http://www.informatics.jax.org/batch   For more information on Gene mutations related to cancer. Atlas of Genetics Oncology: http://atlasgeneticsoncology.org/index.html Section 14.2: For CRISPR-Cas9 systems: If CRISPR-Cas9 is used for other purposes for example, ectopic regulation of gene expression or to deliver cargos to targeted genomic locations, describe the genes that will be regulated or the cargos and the targeted locations.  Are the guide RNA and nuclease on the same plasmid, vector or delivery vehicle? If yes, can this plasmid, vector or delivery vehicle transfer or infect a human cell and can the gRNA or CRISPR nuclease be expressed in human cells? Does the CRISPR research involve viral vector? If yes, a Genome Target Scan (GT-Scan) for off target effects by your gRNA must be complete. This is necessary to determine if there is homology to human DNA and for assessing the risk of potential exposure in the event of an unanticipated incident. An off-target database is available at http://www.rgenome.net/cas-offinder/ Questions? Contact: ibc@uci.edu 949-824-8024