久久这里只有精品8丨久久免费一区丨欧美日韩一区三区丨欧美巨乳在线观看丨欧美久久久久久久久久久丨亚洲成a人片在线播放丨日本少妇翘臀啪啪无遮挡丨青青草原国产av福利网站丨www成人精品免费网站青椒丨又爽又黄又无遮挡的视频丨新婚若妻侵犯中文字幕丨国产区福利丨精品无码av人在线观看丨91看片www丨都市乱淫丨在线|国产精品女主播阳台丨少妇又粗又猛又爽又黄的视频丨欧美精品国产丨蜜桃视频欧美丨岛国片在线播放97丨欧美性天天影院丨亚洲欧美男人天堂丨亚洲欧美日韩国产综合在线一区丨乱老年女人伦免费视频丨制服丝袜亚洲中文综合懂色

 Saturation mutagenesis—coupled to an appropriate biological assay—represents a fundamental means of achieving a high-resolution understanding of regulatory and protein-coding nucleic acid sequences of interest. However, mutagenized sequences introduced in trans on episomes or via random or “safe-harbour” integration fail to capture the native context of the endogenous chromosomal locus. This shortcoming markedly limits the interpretability of the resulting measurements of mutational impact. Here, we couple CRISPR/Cas9 RNA-guided cleavage with multiplex homology-directed repair using a complex library of donor templates to demonstrate saturation editing of genomic regions. In exon 18 of BRCA1, we replac a six-base-pair (bp) genomic region with all possible hexamers, or the full exon with all possible single nucleotide variants (SNVs), and measure strong effects on transcript abundance attributable to nonsense-mediated decay and exonic splicing elements. We similarly perform saturation genome editing of a well-conserved coding region of an essential gene, DBR1, and measure relative effects on growth that correlate with functional impact. Measurement of the functional consequences of large numbers of mutations with saturation genome editing will potentially facilitate high-resolution functional dissection of both cis-regulatory elements and trans-acting factors, as well as the interpretation of variants of uncertain significance observed in clinical sequencing.