A recombineering based approach for high-throughput conditional knockout targeting vector construction

被引:93
作者
Chan, Waiin
Costantino, Nina
Li, Ruixue
Lee, Song Choon
Su, Qin
Melvin, David
Court, Donald L. [1 ]
Liu, Pentao
机构
[1] Wellcome Trust Sanger Inst, Cambridge CB10 1HH, England
[2] NCI Frederick, Ft Detrick, MD 21702 USA
基金
英国惠康基金;
关键词
ESCHERICHIA-COLI; MOUSE GENOME; HOMOLOGOUS RECOMBINATION; BACTERIOPHAGE-LAMBDA; MAMMALIAN-CELLS; DNA; CHROMOSOME; SYSTEM; TRANSCRIPTION; STEP;
D O I
10.1093/nar/gkm163
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Functional analysis of mammalian genes in vivo is primarily achieved through analysing knockout mice. Now that the sequencing of several mammalian genomes has been completed, understanding functions of all the genes represents the next major challenge in the post-genome era. Generation of knockout mutant mice has currently been achieved by many research groups but only by making individual knockouts, one by one. New technological advances and the refinements of existing technologies are critical for genome-wide targeted mutagenesis in the mouse. We describe here new recombineering reagents and protocols that enable recombineering to be carried out in a 96-well format. Consequently, we are able to construct 96 conditional knockout targeting vectors simultaneously. Our new recombineering system makes it a reality to generate large numbers of precisely engineered DNA constructs for functional genomics studies.
引用
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页数:13
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共 36 条
  • [1] A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction
    Adams, DJ
    Quail, MA
    Cox, T
    van der Weyden, L
    Gorick, BD
    Su, Q
    Chan, WI
    Davies, R
    Bonfield, JK
    Law, F
    Humphray, S
    Plumb, B
    Liu, PT
    Rogers, J
    Bradley, A
    [J]. GENOMICS, 2005, 86 (06) : 753 - 758
  • [2] The connection between transcription and genomic instability
    Aguilera, A
    [J]. EMBO JOURNAL, 2002, 21 (03) : 195 - 201
  • [3] ANGRAND PO, 1999, NUCLEIC ACIDS RES, V27, P16
  • [4] The knockout mouse project
    Austin, CP
    Battey, JF
    Bradley, A
    Bucan, M
    Capecchi, M
    Collins, FS
    Dove, WF
    Duyk, G
    Dymecki, S
    Eppig, JT
    Grieder, FB
    Heintz, N
    Hicks, G
    Insel, TR
    Joyner, A
    Koller, BH
    Lloyd, KCK
    Magnuson, T
    Moore, MW
    Nagy, A
    Pollock, JD
    Roses, AD
    Sands, AT
    Seed, B
    Skarnes, WC
    Snoddy, J
    Soriano, P
    Stewart, DJ
    Stewart, F
    Stillman, B
    Varmus, H
    Varticovski, L
    Verma, IM
    Vogt, TF
    von Melchner, H
    Witkowski, J
    Woychik, RP
    Wurst, W
    Yancopoulos, GD
    Young, SG
    Zambrowicz, B
    [J]. NATURE GENETICS, 2004, 36 (09) : 921 - 924
  • [5] The European dimension for the mouse genome mutagenesis program
    Auwerx, J
    Avner, P
    Baldock, R
    Ballabio, A
    Balling, R
    Barbacid, M
    Berns, A
    Bradley, A
    Brown, S
    Carmeliet, P
    Chambon, P
    Cox, R
    Davidson, D
    Davies, K
    Duboule, D
    Forejt, J
    Granucci, F
    Hastie, N
    de Angelis, MH
    Jackson, I
    Kioussis, D
    Kollias, G
    Lathrop, M
    Lendahl, U
    Malumbres, M
    von Melchner, H
    Müller, W
    Partanen, J
    Ricciardi-Castagnoli, P
    Rigby, P
    Rosen, B
    Rosenthal, N
    Skarnes, B
    Stewart, AF
    Thornton, J
    Tocchini-Valentini, G
    Wagner, E
    Wahli, W
    Wurst, W
    [J]. NATURE GENETICS, 2004, 36 (09) : 925 - 927
  • [6] SENSITIVE MUTANTS OF BACTERIOPHAGE LAMBDA
    CAMPBELL, A
    [J]. VIROLOGY, 1961, 14 (01) : 22 - &
  • [7] COMPARATIVE MOLECULAR-BIOLOGY OF LAMBDOID PHAGES
    CAMPBELL, A
    [J]. ANNUAL REVIEW OF MICROBIOLOGY, 1994, 48 : 193 - 222
  • [8] A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
    Cotta-De-Almeida, V
    Schonhoff, S
    Shibata, T
    Leiter, A
    Snapper, SB
    [J]. GENOME RESEARCH, 2003, 13 (09) : 2190 - 2194
  • [9] Mini-λ:: a tractable system for chromosome and BAC engineering
    Court, DL
    Swaminathan, S
    Yu, DG
    Wilson, H
    Baker, T
    Bubunenko, M
    Sawitzke, J
    Sharan, SK
    [J]. GENE, 2003, 315 : 63 - 69
  • [10] Genetic engineering using homologous recombination
    Court, DL
    Sawitzke, JA
    Thomason, LC
    [J]. ANNUAL REVIEW OF GENETICS, 2002, 36 : 361 - 388