Chari Raj, Mali Prashant, Moosburner Mark, Church George M
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Department of Bioengineering, University of California, San Diego, La Jolla, California, USA.
Nat Methods. 2015 Sep;12(9):823-6. doi: 10.1038/nmeth.3473. Epub 2015 Jul 13.
We developed an in vivo library-on-library methodology to simultaneously assess single guide RNA (sgRNA) activity across ∼1,400 genomic loci. Assaying across multiple human cell types and end-processing enzymes as well as two Cas9 orthologs, we unraveled underlying nucleotide sequence and epigenetic parameters. Our results and software (http://crispr.med.harvard.edu/sgRNAScorer) enable improved design of reagents, shed light on mechanisms of genome targeting, and provide a generalizable framework to study nucleic acid-nucleic acid interactions and biochemistry in high throughput.
我们开发了一种体内文库对文库方法，以同时评估跨越约1400个基因组位点的单向导RNA（sgRNA）活性。通过在多种人类细胞类型、末端加工酶以及两种Cas9直系同源物中进行检测，我们揭示了潜在的核苷酸序列和表观遗传参数。我们的研究结果和软件（http://crispr.med.harvard.edu/sgRNAScorer）能够改进试剂设计，阐明基因组靶向机制，并提供一个可推广的框架，用于高通量研究核酸 - 核酸相互作用和生物化学。
