内生
计算生物学
功能(生物学)
吞吐量
蛋白质功能
计算机科学
生物
遗传学
基因
生物化学
电信
无线
作者
Joonwon Kim,Anton Kratz,Shiye Chen,James J. Sheng,Hark Kyun Kim,Liudeng Zhang,Brijesh Kumar Singh,Alejandro Chavez
出处
期刊:Science Advances
[American Association for the Advancement of Science (AAAS)]
日期:2024-05-03
卷期号:10 (18)
标识
DOI:10.1126/sciadv.adg8771
摘要
To facilitate the interrogation of protein function at scale, we have developed high-throughput insertion of tags across the genome (HITAG). HITAG enables users to rapidly produce libraries of cells, each with a different protein of interest C-terminally tagged. HITAG is based on a modified strategy for performing Cas9-based targeted insertions, coupled with an improved approach for selecting properly tagged lines. Analysis of the resulting clones generated by HITAG reveals high tagging specificity, with most successful tagging events being indel free. Using HITAG, we fuse mCherry to a set of 167 stress granule–associated proteins and elucidate the features that drive a subset of proteins to strongly accumulate within these transient RNA-protein granules.
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