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Nucleic Acids Res. 2017 Jun 20;45(11):e101. doi: 10.1093/nar/gkx181.

Flexible CRISPR library construction using parallel oligonucleotide retrieval.

Author information

1
Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
2
Thoracic and Gastrointestinal Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
3
Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.

Abstract

CRISPR/Cas9-based gene knockout libraries have emerged as a powerful tool for functional screens. We present here a set of pre-designed human and mouse sgRNA sequences that are optimized for both high on-target potency and low off-target effect. To maximize the chance of target gene inactivation, sgRNAs were curated to target both 5΄ constitutive exons and exons that encode conserved protein domains. We describe here a robust and cost-effective method to construct multiple small sized CRISPR library from a single oligo pool generated by array synthesis using parallel oligonucleotide retrieval. Together, these resources provide a convenient means for individual labs to generate customized CRISPR libraries of variable size and coverage depth for functional genomics application.

PMID:
28334828
PMCID:
PMC5499874
DOI:
10.1093/nar/gkx181
[Indexed for MEDLINE]
Free PMC Article

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