Element Coding Based Accurate Evaluation of CRISPR/Cas9 Initial Cleavage

Chemical Science ◽

10.1039/d1sc03599a ◽

2021 ◽

Author(s):

Jianyu Hu ◽

Rui Liu ◽

Jing Zhou ◽

Yi Lv

Keyword(s):

Gene Editing ◽

Kinetic Mechanism ◽

Accurate Evaluation ◽

Initial Cleavage

As a powerful gene editing tool, the kinetic mechanism of CRISPR/Cas9 has been the focus to its further application. Initial cleavage events as the first domino followed by nuclease end...

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CrispRVariants: precisely charting the mutation spectrum in genome engineering experiments

10.1101/034140 ◽

2015 ◽

Cited By ~ 1

Author(s):

Helen Lindsay ◽

Alexa Burger ◽

Berthin Biyong ◽

Anastasia Felker ◽

Christopher Hess ◽

...

Keyword(s):

Genomic Dna ◽

Gene Editing ◽

Genome Engineering ◽

Careful Consideration ◽

Functional Screening ◽

Sequencing Data ◽

Web Tool ◽

Therapeutic Gene ◽

Accurate Evaluation ◽

Mutagenic Process

CRISPR-Cas9 and related technologies efficiently alter genomic DNA at targeted positions and have far-reaching implications for functional screening and therapeutic gene editing. Understanding and unlocking this potential requires accurate evaluation of editing efficiency. We show that methodological decisions for analyzing sequencing data can significantly affect mutagenesis efficiency estimates and we provide a comprehensive R-based toolkit, CrispRVariants and accompanying web tool CrispRVariantsLite, that resolves and localizes individual mutant alleles with respect to the endonuclease cut site. CrispRVariants-enabled analyses of newly generated and existing genome editing datasets underscore how careful consideration of the full variant spectrum gives insight toward effective guide and amplicon design as well as the mutagenic process.

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