A conditional knockout resource for the genome-wide study of mouse gene function

被引：1275

作者：

Skarnes, William C. ^{[1
]}

Rosen, Barry ^{[1
]}

West, Anthony P. ^{[1
]}

Koutsourakis, Manousos ^{[1
]}

Bushell, Wendy ^{[1
]}

Iyer, Vivek ^{[1
]}

Mujica, Alejandro O. ^{[1
]}

Thomas, Mark ^{[1
]}

Harrow, Jennifer ^{[1
]}

Cox, Tony ^{[1
]}

Jackson, David ^{[1
]}

Severin, Jessica ^{[1
]}

Biggs, Patrick ^{[1
]}

Fu, Jun ^{[2
]}

Nefedov, Michael ^{[3
]}

de Jong, Pieter J. ^{[3
]}

Stewart, A. Francis ^{[2
]}

Bradley, Allan ^{[1
]}

机构：

[1] Wellcome Trust Sanger Inst, Cambridge CB10 1SA, England

[2] Tech Univ Dresden, Biotechnol Zentrum, D-01062 Dresden, Germany

[3] Childrens Hosp, Oakland Res Inst, Oakland, CA 94609 USA

来源：

NATURE | 2011年 / 474卷 / 7351期

基金：

美国国家卫生研究院; 英国惠康基金;

关键词：

EMBRYONIC STEM-CELLS; HOMOLOGOUS RECOMBINATION; CAENORHABDITIS-ELEGANS; NEGATIVE SELECTION; RAT BLASTOCYSTS; EXPRESSION; TRAP; MICE; REPORTER; VECTOR;

D O I：

10.1038/nature10163

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Gene targeting in embryonic stem cells has become the principal technology for manipulation of the mouse genome, offering unrivalled accuracy in allele design and access to conditional mutagenesis. To bring these advantages to the wider research community, large-scale mouse knockout programmes are producing a permanent resource of targeted mutations in all protein-coding genes. Here we report the establishment of a high-throughput gene-targeting pipeline for the generation of reporter-tagged, conditional alleles. Computational allele design, 96-well modular vector construction and high-efficiency gene-targeting strategies have been combined to mutate genes on an unprecedented scale. So far, more than 12,000 vectors and 9,000 conditional targeted alleles have been produced in highly germline-competent C57BL/6N embryonic stem cells. High-throughput genome engineering highlighted by this study is broadly applicable to rat and human stem cells and provides a foundation for future genome-wide efforts aimed at deciphering the function of all genes encoded by the mammalian genome.

引用

页码：337 / U361

页数：8

共 48 条

[21] DISRUPTION OF THE PROTO-ONCOGENE INT-2 IN MOUSE EMBRYO-DERIVED STEM-CELLS - A GENERAL STRATEGY FOR TARGETING MUTATIONS TO NON-SELECTABLE GENES [J].

MANSOUR, SL ;

THOMAS, KR ;

CAPECCHI, MR .

NATURE, 1988, 336 (6197) :348-352

[22] Functional analysis of secreted and transmembrane proteins critical to mouse development [J].

Mitchell, KJ ;

Pinson, KI ;

Kelly, OG ;

Brennan, J ;

Zupicich, L ;

Scherz, P ;

Leighton, PA ;

Goodrich, LV ;

Lu, XW ;

Avery, BJ ;

Tate, P ;

Dill, K ;

Pangilinan, E ;

Wakenight, P ;

Tessier-Lavigne, M ;

Skarnes, WC .

NATURE GENETICS, 2001, 28 (03) :241-249

[23]

Osoegawa K, 2000, GENOME RES, V10, P116

[24] Agouti C57BL/6N embryonic stem cells for mouse genetic resources [J].

Pettitt, Stephen J. ;

Liang, Qi ;

Rairdan, Xin Y. ;

Moran, Jennifer L. ;

Prosser, Haydn M. ;

Beier, David R. ;

Lloyd, Kent C. ;

Bradley, Allan ;

Skarnes, William C. .

NATURE METHODS, 2009, 6 (07) :493-U34

[25]

Poser I, 2008, NAT METHODS, V5, P409, DOI [10.1038/nmeth.1199, 10.1038/NMETH.1199]

[26] FO generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses [J].

Poueymirou, William T. ;

Auerbach, Wojtek ;

Frendewey, David ;

Hickey, Joseph F. ;

Escaravage, Jennifer M. ;

Esau, Lakeisha ;

Dore, Anthony T. ;

Stevens, Sean ;

Adams, Niels C. ;

Dominguez, Melissa G. ;

Gale, Nicholas W. ;

Yancopoulos, Geore D. ;

DeChiara, Thomas M. ;

Valenzuela, David M. .

NATURE BIOTECHNOLOGY, 2007, 25 (01) :91-99

[27] HIGHLY EFFICIENT GENE TARGETING IN EMBRYONIC STEM-CELLS THROUGH HOMOLOGOUS RECOMBINATION WITH ISOGENIC DNA CONSTRUCTS [J].

RIELE, HT ;

MAANDAG, ER ;

BERNS, A .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (11) :5128-5132

[28] The IKMC web portal: a central point of entry to data and resources from the International Knockout Mouse Consortium [J].

Ringwald, Martin ;

Iyer, Vivek ;

Mason, Jeremy C. ;

Stone, Kevin R. ;

Tadepally, Hamsa D. ;

Kadin, James A. ;

Bult, Carol J. ;

Eppig, Janan T. ;

Oakley, Darren J. ;

Briois, Sebastien ;

Stupka, Elia ;

Maselli, Vincenza ;

Smedley, Damian ;

Liu, Songyan ;

Hansen, Jens ;

Baldock, Richard ;

Hicks, Geoff G. ;

Skarnes, William C. .

NUCLEIC ACIDS RESEARCH, 2011, 39 :D849-D855

[29] GERM-LINE TRANSMISSION OF GENES INTRODUCED INTO CULTURED PLURIPOTENTIAL CELLS BY RETROVIRAL VECTOR [J].

ROBERTSON, E ;

BRADLEY, A ;

KUEHN, M ;

EVANS, M .

NATURE, 1986, 323 (6087) :445-448

[30] Towards a proteome-scale map of the human protein-protein interaction network [J].

Rual, JF ;

Venkatesan, K ;

Hao, T ;

Hirozane-Kishikawa, T ;

Dricot, A ;

Li, N ;

Berriz, GF ;

Gibbons, FD ;

Dreze, M ;

Ayivi-Guedehoussou, N ;

Klitgord, N ;

Simon, C ;

Boxem, M ;

Milstein, S ;

Rosenberg, J ;

Goldberg, DS ;

Zhang, LV ;

Wong, SL ;

Franklin, G ;

Li, SM ;

Albala, JS ;

Lim, JH ;

Fraughton, C ;

Llamosas, E ;

Cevik, S ;

Bex, C ;

Lamesch, P ;

Sikorski, RS ;

Vandenhaute, J ;

Zoghbi, HY ;

Smolyar, A ;

Bosak, S ;

Sequerra, R ;

Doucette-Stamm, L ;

Cusick, ME ;

Hill, DE ;

Roth, FP ;

Vidal, M .

NATURE, 2005, 437 (7062) :1173-1178

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