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An improved strategy for CRISPR/Cas9 gene knockout and subsequent wildtype and mutant gene rescue

13 Feb 2020

Abstract excerpt

A fluorescence marker mOrange was inserted to the popular pLentiCrispr-V2 to create pLentiCrispr-V2-mOrange (V2mO) that contained both a puromycin selection and a fluorescent marker, making viral production and target transduction visible. Lentiviruses packaged with this plasmid and appropriate guide RNAs (gRNAs) successfully knocked out the genes RhoA, Gli1, and Gal3 in human gastric cancer cell lines. Cas9-gRNA...

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PMID: 32053639
DOI: 10.1371/journal.pone.0228910

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