AI Article Synopsis
- CRISPR/Cas9 gene knockout libraries offer a powerful approach for conducting functional screens by providing specific sgRNA sequences optimized for effectiveness and minimal off-target effects.
- These sgRNAs specifically target important regions of genes, including 5' exons and conserved protein domains, to enhance the likelihood of successfully inactivating target genes.
- The described method allows labs to easily create custom CRISPR libraries of different sizes and coverage from a single oligo pool, making it resource-efficient for functional genomics research.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499874 | PMC |
| http://dx.doi.org/10.1093/nar/gkx181 | DOI Listing |
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