Flexible CRISPR library construction using parallel oligonucleotide retrieval.
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.
Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Abigail
A
Read
Thoracic and Gastrointestinal Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Shaojian
S
Gao
Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Eric
E
Batchelor
Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Ji
J
Luo
D000818:Animals; D064113:CRISPR-Cas Systems; D005091:Exons; D055786:Gene Knockout Techniques; D015723:Gene Library; D006801:Humans; D051379:Mice; D009841:Oligonucleotides / Q000235:genetics*
D009841:Oligonucleotides