Xi Dawn Chen, Zeyu Chen, George Wythes, Yifan Zhang, Benno C. Orr, Gary Sun, Yu-Kai Chao, Andrea Navarro Torres, Ka Thao, Mounica Vallurupalli, Jing Sun, Mehdi Borji, Emre Tkacik, Haiqi Chen, Bradley E. Bernstein, Fei Chen
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引用次数: 0
Abstract
Deciphering the context-specific relationship between sequence and function is a major challenge in genomics. Existing tools for inducing locus-specific hypermutation and evolution in the native genome context are limited. Here we present a programmable platform for long-range, locus-specific hypermutation called helicase-assisted continuous editing (HACE). HACE leverages CRISPR-Cas9 to target a processive helicase-deaminase fusion that incurs mutations across large (>1000–base pair) genomic intervals. We applied HACE to identify mutations in mitogen-activated protein kinase kinase 1 (MEK1) that confer kinase inhibitor resistance, to dissect the impact of individual variants in splicing factor 3B subunit 1 (SF3B1)–dependent missplicing, and to evaluate noncoding variants in a stimulation-dependent immune enhancer of CD69. HACE provides a powerful tool for investigating coding and noncoding variants, uncovering combinatorial sequence-to-function relationships, and evolving new biological functions.
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