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Article

Quantitative assessment of engineered Cas9 variants for target specificity enhancement by single-molecule reaction pathway analysis.

Nucleic acids research - 8 Nov 2021

Bak So Young, Jung Youngri, Park Jinho, Sung Keewon, Jang Hyeon-Ki, Bae Sangsu, Kim Seong Keun

Abstract excerpt

There have been many engineered Cas9 variants that were developed to minimize unintended cleavage of off-target DNAs, but detailed mechanism for the way they regulate the target specificity through DNA:RNA heteroduplexation remains poorly understood. We used single-molecule FRET assay to follow the dynamics of DNA:RNA heteroduplexation for various engineered Cas9 variants with respect to on-target and off-target...

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PMID: 34554247
DOI: 10.1093/nar/gkab858
PMCID: PMC8565309

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