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European Biopharmaceutical Review

Choosing the Right CRISPR Tools to Generate Your Genetically Modified Mice

Genetically modified mice have proven to be an invaluable tool in the biotechnology and pharmaceutical research arsenal, serving as models for human disease and drug development. As a testament to the importance of this tool, the 2007 Nobel Prize in Physiology or Medicine was awarded jointly to Dr Capecchi, Dr Evans, and Dr Smithies for their work on the generation of transgenic mice (1).

The generation of genetically modified mice is incredibly vital to life science research, so much so that it has persisted despite the laborious process required to generate them. The traditional embryonic stem (ES) cell method can take anywhere from eight months to two years (at considerable cost), before obtaining edited mice suitable for downstream experiments. This lengthy process is due to the large – and often manual – number of steps involved, including large donor plasmid generation, transfection, selection, and expansion of ES cells (2). The traditional ES cell method, which relies upon spontaneous integration of a donor construct, yields ~1% integration, even with 10kb homology arms (3). Therefore, a more efficient approach for integrating DNA templates is needed to speed up the overall process of generating genetically modified mice.

CRISPR Has Simplified Gene Editing In Mice

The discovery of CRISPR has made gene editing accessible to all researchers. While many CRISPR systems, from both bacteria and archaea, have been characterised, the CRISPR-Cas9 system has established itself as the dominant gene editing tool for researchers. The CRISPR-Cas9 system utilises a Cas9 endonuclease and a guide RNA, which is complementary to genomic region of interest; the guide RNA will direct the Cas9 protein to a specific genomic target, where a double strand break is generated. CRISPR can target most genomic loci and the targeting of a new specific loci requires changing of only the protospacer sequence (~20nt); this allows for target-specific editing tools to be ready days after the need arises.

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James Chon, PhD, is a Field Application Scientist at GenScript USA, where he provides technical support and designs project proposals related to the company's CRISPR services. James often provides webinars and presents posters in support of the company’s CRISPR business. Previously at GenScript, he served as a Senior Technical Account Manager for projects in gene and peptide synthesis, CRISPR services, protein purification and library generation. James received a Bachelor's degree in Biology from The City College of New York, US, and a doctorate in molecular and cellular biology from Cornell University, US.
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James Chon
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