Vertical Integration of CRISPR/Cas9 Genome Engineering in Drug Discovery

Once in a while discoveries are made that greatly impact the world. In the biology field, the CRISPR/Cas9 system is such a revolutionizing technology that brought rapid, efficient and precise genome engineering in a variety of species within reach of many researchers. The beauty of the system lies in the use of guide RNAs(gRNAs) that direct the Cas9 endonuclease to a specific site in the genome where it induces a double-stranded break. The repair of this break allows a gene editing event either via the non-homologous end joining (NHEJ) or homology-driven repair (HDR) pathway.

Since its discovery as a genome engineering tool in eukaryotes in 2013, the research field has embraced this technology for different applications ranging from basic research into gene-function relationships to the potential use for gene therapy. In this webinar, the speaker will focus on the utilization of CRISPR/Cas9 technology in drug discovery, with an emphasis on generation of in vitro models for high-throughput screening, and creating new mouse models.

Several CRISPR/Cas9 case studies will be discussed during this webinar covering different aspects of the drug discovery process. One example is the integration of a transgene into a safe harbor locus of the HEK293 cell line using a single-stranded oligo donor as repair template. Other case studies focus on the generation of knock-out or single nucleotide polymorphism (SNP)-corrected mice by CRISPR/Cas9 gene editing of mouse embryonic stem cells (mESC) using the NHEJ and the HDR repair pathway and subsequent injections of the targeted mESC into blastocysts. CRISPR/Cas9 technology was also applied for gene editing of mouse tumor cell lines to generate syngeneic mouse models matching human tumors, using a two gRNA approach to delete an entire exon for full knock-out of the targeted gene. Future activities of Charles River in the field CRISPR/Cas9 will expand to the use of gRNA libraries for target discovery, similar to the Charles River SilenceSelect^® RNAi interference technology.

Speaker

Anne-Marie Zuurmond, Ph.D., Associate Director of Genome Engineering, Discovery, Charles River Laboratories

Dr. Anne-Marie Zuurmond is the Associate Director of Genome Engineering for Charles River Discovery, based at their laboratory in Leiden, the Netherlands. She received a M.Sc. in Molecular Biology from the Leiden University and did her Ph.D. research on the prokaryotic protein synthesis machinery at the Leiden Institute of Chemistry. Over the past 11 years, Dr. Zuurmond has been active as project leader in a CRO environment managing projects in the field of fibrosis, arthritis, metabolic syndrome, and the central nervous system. The nature of these projects ranged from target discovery using RNA interference, in vitro and in vivo target validation using viral systems for shRNA delivery to mechanistic and compound efficacy studies in animal models of disease. When Dr. Zuurmond joined Charles River in January 2015 she initiated the implementation of the CRISPR/Cas9 technology in the drug discovery and animal research models business of Charles River. Since then, she has used this genome editing tool in creating knock-out and knock-in cell lines for in vitro drug discovery and the generation of novel transgenic mouse models. She envisions gRNA library screening to become an additional tool in target discovery alongside sh/siRNA library screening and that the process from hit discovery, hit-to-lead and lead-to-candidate will benefit from the possibilities that this technology offers in generating in vitro and in vivo models for screening, validation and compound testing.

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