CRISPR-Cas9 Genome Editing in Human Cell Lines with Donor Vector Made by Gibson Assembly

利用 Gibson Assembly 制造的供体载体在人类细胞系中进行 CRISPR-Cas9 基因组编辑

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作者:Nirakar Sahoo, Victoria Cuello, Shreya Udawant, Carl Litif, Julie A Mustard, Megan Keniry

Abstract

CRISPR Cas9 genome editing allows researchers to modify genes in a multitude of ways including to obtain deletions, epitope-tagged loci, and knock-in mutations. Within 6 years of its initial application, CRISPR-Cas9 genome editing has been widely employed, but disadvantages to this method, such as low modification efficiencies and off-target effects, need careful consideration. Obtaining custom donor vectors can also be expensive and time-consuming. This chapter details strategies to overcome barriers to CRISPR-Cas9 genome editing as well as recent developments in employing this technique.

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