Vejnar Charles E, Moreno-Mateos Miguel A, Cifuentes Daniel, Bazzini Ariel A, Giraldez Antonio J
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510 Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut 06520.
Cold Spring Harb Protoc. 2016 Oct 3;2016(10):2016/10/pdb.top090894. doi: 10.1101/pdb.top090894.
The CRISPR-Cas9 system uncovered in bacteria has emerged as a powerful genome-editing technology in eukaryotic cells. It consists of two components-a single guide RNA (sgRNA) that directs the Cas9 endonuclease to a complementary DNA target site. Efficient targeting of individual genes requires highly active sgRNAs. Recent efforts have made significant progress in understanding the sequence features that increase sgRNA activity. In this introduction, we highlight advancements in the field of CRISPR-Cas9 targeting and discuss our web tool CRISPRscan, which predicts the targeting activity of sgRNAs and improves the efficiency of the CRISPR-Cas9 system for in vivo genome engineering.
在细菌中发现的CRISPR-Cas9系统已成为真核细胞中一种强大的基因组编辑技术。它由两个组件组成——一个单向导RNA（sgRNA），可将Cas9核酸内切酶引导至互补的DNA靶位点。高效靶向单个基因需要高活性的sgRNA。最近的研究在了解增加sgRNA活性的序列特征方面取得了重大进展。在本引言中，我们重点介绍CRISPR-Cas9靶向领域的进展，并讨论我们的网络工具CRISPRscan，该工具可预测sgRNA的靶向活性，并提高CRISPR-Cas9系统用于体内基因组工程的效率。
