Synthetic switch to minimize CRISPR off-target effects by self-restricting Cas9 transcription and translation

• Published in: Nucleic acids research Vol. 47; no. 3; p. e13
• Main Authors: Shen, Chih-Che, Hsu, Mu-Nung, Chang, Chin-Wei, Lin, Mei-Wei, Hwu, Jih-Ru, Tu, Yi, Hu, Yu-Chen
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 20.02.2019
• Subjects: Cell Line; CRISPR-Associated Protein 9 - biosynthesis; CRISPR-Associated Protein 9 - genetics; CRISPR-Cas Systems; Gene Editing; Gene Expression Regulation; Humans; Kinetics; Methods Online; Mutagenesis; Mutation; Protein Biosynthesis; Transcription, Genetic

Abstract CRISPR/Cas9 is a powerful genome editing system but uncontrolled Cas9 nuclease expression triggers off-target effects and even in vivo immune responses. Inspired by synthetic biology, here we built a synthetic switch that self-regulates Cas9 expression not only in the transcription step by...