Gene editing has become a firmly established technology within the discovery sciences arena. With the advent of CRISPR/Cas9 systems, the researcher’s ability to find an active nuclease for nearly any region of any genome is now a reality. Even with better nucleases available, those who routinely utilize gene editing tools to manipulate cell lines encounter other significant challenges that pose a barrier to building the “correct” cell line model. As a well-established partner for custom cell line engineering, we at Merck KGaA, Darmstadt, Germany have encountered many of these challenges and have developed and/or implemented a number of methods to circumvent them. A summary of the various formats of CRISPR/Cas9 tools, and recommendations for when to use each will also be presented.
What will you learn?
- Newest strategies for clone screening and genotyping
- Improved workflow for cell line characterization
- Methods to overcome poor transfection efficiency
- Strategies for recalcitrant cell lines
Who may this interest?
- Researchers/scientists; Target discovery, target validation, early oncology discovery, immuno-oncology, immunotherapy, T-cell biology
Principal R&D Scientist, Supervisor, Cell Design Studio
Mark obtained his Ph.D. in Biochemistry and Molecular Biology from Saint Louis University School of Medicine, where he used RNAi in Drosophila models to elucidate developmental and biochemical roles for RNA polymerase II-associated transcription factors. Following graduate school, he was a Postdoctoral Fellow at Washington University in St. Louis, where he investigated signaling pathways involved in the development of human meningioma. Mark joined joined MilliporeSigma* in 2006, and he has worked in the areas of biotherapeutic production, stem cell applications, and gene regulation. He currently supervises the Cell Design Studio team, which performs custom cell line engineering for research and pharma markets using ZFN and CRISPR technologies.
* The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.
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