Genome-scale loss-of-function screening with a lentiviral RNAi library

被引：290

作者：

Root, David E.

Hacohen, Nir

Hahn, William C.

Lander, Eric S.

Sabatini, David M.

机构：

[1] MIT, Broad Inst, Cambridge, MA 02142 USA

[2] Harvard, Cambridge, MA 02142 USA

[3] Massachusetts Gen Hosp, Ctr Immunol & Inflammatory Dis, Boston, MA 02114 USA

[4] Harvard Univ, Sch Med, Boston, MA 02115 USA

[5] Dana Farber Canc Inst, Ctr Canc Genome Discovery, Dept Med Oncol, Boston, MA 02115 USA

[6] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA

[7] MIT, Dept Biol, Cambridge, MA 02139 USA

来源：

NATURE METHODS | 2006年 / 3卷 / 09期

关键词：

D O I：

10.1038/NMETH924

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The discovery that RNA interference (RNAi) is functional in mammalian cells led us to form The RNAi Consortium (TRC) with the goal of enabling large-scale loss-of-function screens through the development of genome-scale RNAi libraries and methodologies for their use. These resources form the basis of a loss-of-function screening platform created at the Broad Institute. Our human and mouse libraries currently contain > 135,000 lentiviral clones targeting 27,000 genes. Initial screening efforts have demonstrated that these libraries and methods are practical and powerful tools for high-throughput lentivirus RNAi screens.

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收藏

页码：715 / 719

页数：5

相关论文

共 23 条

[1] A large-scale RNAi screen in human cells identifies new components of the p53 pathway [J].

Berns, K ;

Hijmans, EM ;

Mullenders, J ;

Brummelkamp, TR ;

Velds, A ;

Heimerikx, M ;

Kerkhoven, RM ;

Madiredjo, M ;

Nijkamp, W ;

Weigelt, B ;

Agami, R ;

Ge, W ;

Cavet, G ;

Linsley, PS ;

Beijersbergen, RL ;

Bernards, R .

NATURE, 2004, 428 (6981) :431-437

[2] 3′ UTR seed matches, but not overall identity, are associated with RNAi off-targets [J].

Birmingham, A ;

Anderson, EM ;

Reynolds, A ;

Ilsley-Tyree, D ;

Leake, D ;

Fedorov, Y ;

Baskerville, S ;

Maksimova, E ;

Robinson, K ;

Karpilow, J ;

Marshall, WS ;

Khvorova, A .

NATURE METHODS, 2006, 3 (03) :199-204

[3] An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors [J].

Brummelkamp, TR ;

Fabius, AWM ;

Mullenders, J ;

Madiredjo, M ;

Velds, A ;

Kerkhoven, RM ;

Bernards, R ;

Beijersbergen, RL .

NATURE CHEMICAL BIOLOGY, 2006, 2 (04) :202-206

[4] Innovation - New tools for functional mammalian cancer genetics [J].

Brummelkamp, TR ;

Bernards, R .

NATURE REVIEWS CANCER, 2003, 3 (10) :781-789

[5] Cancer-associated PP2A Aα subunits induce functional haploinsufficiency and tumorigenicity [J].

Chen, W ;

Arroyo, JD ;

Timmons, JC ;

Possemato, R ;

Hahn, WC .

CANCER RESEARCH, 2005, 65 (18) :8183-8192

[6] A third-generation lentivirus vector with a conditional packaging system [J].

Dull, T ;

Zufferey, R ;

Kelly, M ;

Mandel, RJ ;

Nguyen, M ;

Trono, D ;

Naldini, L .

JOURNAL OF VIROLOGY, 1998, 72 (11) :8463-8471

[7] Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells [J].

Elbashir, SM ;

Harborth, J ;

Lendeckel, W ;

Yalcin, A ;

Weber, K ;

Tuschl, T .

NATURE, 2001, 411 (6836) :494-498

[8] Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences [J].

Follenzi, A ;

Ailles, LE ;

Bakovic, S ;

Geuna, M ;

Naldini, L .

NATURE GENETICS, 2000, 25 (02) :217-+

[9] Unlocking the potential of the human genome with RNA interference [J].

Hannon, GJ ;

Rossi, JJ .

NATURE, 2004, 431 (7006) :371-378

[10] Expression profiling reveals off-target gene regulation by RNAi [J].

Jackson, AL ;

Bartz, SR ;

Schelter, J ;

Kobayashi, SV ;

Burchard, J ;

Mao, M ;

Li, B ;

Cavet, G ;

Linsley, PS .

NATURE BIOTECHNOLOGY, 2003, 21 (06) :635-637