Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA.
MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston, Houston, TX, 77030, USA.
Nat Commun. 2019 Jul 17;10(1):3144. doi: 10.1038/s41467-019-10963-x.
Capitalizing on the inherent multiplexing capability of AsCpf1, we developed a multiplexed, high-throughput screening strategy that minimizes library size without sacrificing gene targeting efficiency. We demonstrated that AsCpf1 can be used for functional genomics screenings and that an AsCpf1-based multiplexed library performs similarly as compared to currently available monocistronic CRISPR/Cas9 libraries, with only one vector required for each gene. We construct the smallest whole-genome CRISPR knock-out library, Mini-human, for the human genome (n = 17,032 constructs targeting 16,977 protein-coding genes), which performs favorably compared to conventional Cas9 libraries.
利用 AsCpf1 的固有多重功能，我们开发了一种多重、高通量筛选策略，在不牺牲基因靶向效率的情况下最小化文库规模。我们证明了 AsCpf1 可用于功能基因组筛选，并且基于 AsCpf1 的多重文库的性能与当前可用的单顺反子 CRISPR/Cas9 文库相当，每个基因仅需要一个载体。我们构建了最小的全基因组 CRISPR 敲除文库 Mini-human，用于人类基因组（针对 16977 个编码蛋白的基因有 17032 个构建物），与传统的 Cas9 文库相比，表现良好。
