Dense sgRNA Library Construction Using a Molecular Chipper Approach.
Dense sgRNA Library Construction Using a Molecular Chipper Approach.
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DOI：
10.21769/bioprotoc.2373
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发表时间：
2017-06
期刊：
影响因子：
0.8
通讯作者：
Jijun Cheng;Wen Pan;Jun Lu
中科院分区：
文献类型：
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作者：
Jijun Cheng;Wen Pan;Jun Lu
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.
