An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology.
An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology.
复制标题
用于酿酒酵母生物学遗传询问的诱导型CRISPR干扰文库。
DOI：
10.1038/s42003-020-01452-9
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发表时间：
2020-11-27
影响因子：
5.9
通讯作者：
Tavazoie S
中科院分区：
文献类型：
--
作者：
Momen-Roknabadi A;Oikonomou P;Zegans M;Tavazoie S
基因组规模的CRISPR干扰（CRISPRi）被广泛用于研究各种生物的细胞过程。尽管酿酒酵母（Saccharomyces cerevisiae）作为一种模式真核生物占据主导地位，但酵母中可诱导的全基因组CRISPRi文库尚未提出。在这里，我们提出了一个全基因组的、可诱导的CRISPRi文库，该文库基于针对酿酒葡萄球菌优化的间隔设计规则。我们已经验证了该文库在各种应用中的基因功能全基因组调查，包括准确发现单倍不足基因和鉴定参与腺嘌呤和精氨酸生物合成的酶和调控基因。该文库的综合性也揭示了转录抑制的精确间隔设计参数，包括位置、核小体占用和核苷酸特征。使用该文库的CRISPRi筛选可以在各种实验条件下以高精度和低错误发现率识别基因和途径，从而能够快速可靠地评估酿酒葡萄球菌的遗传功能和相互作用。Momen-Roknabadi及其同事提出了一个可诱导的酵母全基因组CRISPRi文库。该文库将实现酵母基因组的全面转录扰动，促进更直接的探索这种模式生物的功能生物学。
Genome-scale CRISPR interference (CRISPRi) is widely utilized to study cellular processes in a variety of organisms. Despite the dominance of Saccharomyces cerevisiae as a model eukaryote, an inducible genome-wide CRISPRi library in yeast has not yet been presented. Here, we present a genome-wide, inducible CRISPRi library, based on spacer design rules optimized for S. cerevisiae. We have validated this library for genome-wide interrogation of gene function across a variety of applications, including accurate discovery of haploinsufficient genes and identification of enzymatic and regulatory genes involved in adenine and arginine biosynthesis. The comprehensive nature of the library also revealed refined spacer design parameters for transcriptional repression, including location, nucleosome occupancy and nucleotide features. CRISPRi screens using this library can identify genes and pathways with high precision and a low false discovery rate across a variety of experimental conditions, enabling rapid and reliable assessment of genetic function and interactions in S. cerevisiae. Momen-Roknabadi and colleagues present an inducible genome-wide CRISPRi library for yeast. This library will enable comprehensive transcriptional perturbations of the yeast genome, facilitating more directed exploration of this model organism’s functional biology.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
影响因子：
4.5
作者：
Girgis, Hany S.;Liu, Yirchung;Ryu, William S.;Tavazoie, Saeed
通讯作者：
Tavazoie, Saeed
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
CRISPR-CAS9介导的基因失活的高度活性SGRNA的合理设计。
影响因子：
46.9
作者：
Doench, John G.;Hartenian, Ella;Graham, Daniel B.;Tothova, Zuzana;Hegde, Mudra;Smith, Ian;Sullender, Meagan;Ebert, Benjamin L.;Xavier, Ramnik J.;Root, David E.
通讯作者：
Root, David E.
影响因子：
16
作者：
Goodarzi H;Elemento O;Tavazoie S
通讯作者：
Tavazoie S