Abstract
It has been challenging for efficient gene editing in cells which are hard to transfect, such as stem cells and primary T cells. Typical sgRNA and Cas9 intracellular delivery techniques are limited by their reliance on cell type and exogenous materials as well as their toxic effects on cells (for example, electroporation) or packaging capacity limitation by viral vectors. Here we established an effective approach for gene editing in different cell types and achieved successful genome editing with CRISPR RNP electroporation. Also, high efficient double knockout mutations could be obtained by electroporation of multiple sgRNAs at one shot. Furthermore, this method also could reduce the exposure time to Cas9 nuclease which may cause potential off-target effects. With the advantages of broad applicability across different cell types, particularly hard-to-transfect cells, and flexibility of application, these methods could potentially enable the development of the stem cell or T cell based therapies.
Citation Format: Peixue Li, Zhao Li, Qing Lin. CRISPR/Cas9 RNP mediated efficient gene editing in hard-to-transfect cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6303.
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