Genome-scale deletion screening of human long non-coding RNAs using a paired-guide RNA CRISPR-Cas9 library

被引：326

作者：

Zhu, Shiyou ^{[1
,2
]}

Li, Wei ^{[3
,4
]}

Liu, Jingze ^{[1
,2
]}

Chen, Chen-Hao ^{[3
,4
]}

Liao, Qi ^{[5
]}

Xu, Ping ^{[1
]}

Xu, Han ^{[6
]}

Xiao, Tengfei ^{[4
,7
]}

Cao, Zhongzheng ^{[1
,8
]}

Peng, Jingyu ^{[1
]}

Yuan, Pengfei ^{[1
]}

Brown, Myles ^{[4
,7
,9
,10
]}

Liu, Xiaole Shirley ^{[3
,4
]}

Wei, Wensheng ^{[1
]}

机构：

[1] Peking Univ, State Key Lab Prot & Plant Gene Res, Sch Life Sci,Biodynam Opt Imaging Ctr BIOPIC, Beijing Adv Innovat Ctr Genom,Peking Tsinghua Ctr, Beijing, Peoples R China

[2] Peking Univ, Peking Univ Tsinghua Univ Natl Inst Biol Sci Join, Beijing, Peoples R China

[3] Harvard TH Chan Sch Publ Hlth, Dana Farber Canc Inst, Dept Biostat & Computat Biol, Boston, MA 02115 USA

[4] Dana Farber Canc Inst, Ctr Funct Canc Epigenet, Boston, MA 02115 USA

[5] Ningbo Univ, Sch Med, Dept Prevent Med, Ningbo, Zhejiang, Peoples R China

[6] Broad Inst MIT & Harvard, Cambridge Ctr, Cambridge, MA USA

[7] Dana Farber Canc Inst, Dept Med Oncol, Div Mol & Cellular Oncol, Boston, MA USA

[8] Peking Univ, Acad Adv Interdisciplinary Studies, Beijing, Peoples R China

[9] Brigham & Womens Hosp, Dept Med, 75 Francis St, Boston, MA 02115 USA

[10] Harvard Med Sch, Boston, MA USA

来源：

NATURE BIOTECHNOLOGY | 2016年 / 34卷 / 12期

基金：

美国国家科学基金会;

关键词：

HUMAN-CELLS; FUNCTIONAL GENOMICS; CRISPR/CAS9; LIBRARY; ESSENTIAL GENES; ACTIVATION; SYSTEM; DNA; IDENTIFICATION; REPRESSION; CANCER;

D O I：

10.1038/nbt.3715

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

CRISPR-Cas9 screens have been widely adopted to analyze coding-gene functions, but high-throughput screening of non-coding elements using this method is more challenging because indels caused by a single cut in non-coding regions are unlikely to produce a functional knockout. A high-throughput method to produce deletions of non-coding DNA is needed. We report a high throughput genomic deletion strategy to screen for functional long non-coding RNAs (IncRNAs) that is based on a lentiviral paired-guide RNA (pgRNA) library. Applying our screening method, we identified 51 IncRNAs that can positively or negatively regulate human cancer cell growth. We validated 9 of 51 IncRNA hits using CRISPR-Cas9-mediated genomic deletion, functional rescue, CRISPR activation or inhibition and gene-expression profiling. Our high-throughput pgRNA genome deletion method will enable rapid identification of functional mammalian non-coding elements.

引用

页码：1279 / 1286

页数：8

共 22 条

[21] Synergistic Antitumor Effect on Bladder Cancer by Rational Combination of Programmed Cell Death 1 Blockade and CRISPR-Cas9-Mediated Long Non-Coding RNA Urothelial Carcinoma Associated 1 Knockout
Zhen, Shuai
Lu, Jiaojiao
Chen, Wei
Zhao, Le
Li, Xu
HUMAN GENE THERAPY, 2018, 29 (12) : 1352 - 1363
[22] Genome-wide screening for novel, drought stress-responsive long non-coding RNAs in drought-stressed leaf transcriptome of drought-tolerant and -susceptible banana (Musa spp) cultivars using Illumina high-throughput sequencing
Muthusamy, M.
Uma, S.
Backiyarani, S.
Saraswathi, M. S.
PLANT BIOTECHNOLOGY REPORTS, 2015, 9 (05) : 279 - 286

← 1 2 3 →