Aug 25, 2017

Dense sgRNA Library Construction Using a Molecular Chipper Approach

Bio-protocol
Jijun ChengJun Lu

Abstract

Genetic screens using single-guide-RNA (sgRNA) libraries and CRISPR technology have been powerful to identify genetic regulators for both coding and noncoding regions of the genome. Interrogating functional elements in noncoding regions requires sgRNA libraries that are densely covering, and ideally inexpensive, easy to implement and flexible for customization. We present a Molecular Chipper protocol for generating dense sgRNA libraries from genomic regions of interest. This approach utilizes a combination of random fragmentation and a Type III restriction enzyme to derive a dense coverage of sgRNA library from input DNA.

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Mentioned in this Paper

Abnormal Fragmented Structure
Type III Site-specific Deoxyribonuclease Activity
Exons
Genome
CRISPR-Cas Systems
Genetic Screening (Procedure)
Genome Assembly Sequence
Genomics
Clustered Regularly Interspaced Short Palindromic Repeats
Type III site-specific deoxyribonuclease

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