An Improved Flp Deleter Mouse in C57Bl/6 Based on Flpo Recombinase

被引:108
作者
Kranz, Andrea
Fu, Jun
Duerschke, Kristin
Weidlich, Stefanie [1 ]
Naumann, Ronald [2 ]
Stewart, A. Francis [1 ]
Anastassiadis, Konstantinos [1 ]
机构
[1] Tech Univ Dresden, BioInnovat Zentrum, Ctr Regenerat Therapies Dresden, D-01307 Dresden, Germany
[2] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
关键词
site specific recombination; conditional mutagenesis; genome engineering; allele design; SITE-SPECIFIC RECOMBINATION; EMBRYONIC STEM-CELLS; MAMMALIAN-CELLS; ESCHERICHIA-COLI; MICE; CRE;
D O I
10.1002/dvg.20641
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Recently, a codon improved version of the Flpe site specific recombinase, termed Flpo, was reported as having greatly improved performance in mammalian cell applications. However, the degree of improvement could not be estimated because essentially no Flpe activity was observed. Here, we compare Flpe and Flpo accurately in a mammalian cell assay to estimate that Flpo is about five times more active than Flpe and similar to Cre and Dre. Consequently, we generated a Flpo deleter mouse line from the JM8 C57BI/6 ES cells used in the EUCOMM and KOMP systematic knock-out programs. In breeding experiments, we show that the Flpo deleter delivers complete recombination using alleles that are incompletely recombined by a commonly used Flpe deleter. This indicates that the Flpo deleter is more efficient. genesis 48:512-520, 2010. (C) 2010 Wiley-Liss, Inc.
引用
收藏
页码:512 / 520
页数:9
相关论文
共 20 条
[11]   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
[12]   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
[13]   High-Efficiency FLP and ΦC31 Site-Specific Recombination in Mammalian Cells [J].
Raymond, Christopher S. ;
Soriano, Philippe .
PLOS ONE, 2007, 2 (01)
[14]   The Kw recombinase, an integrase from Kluyveromyces waltii [J].
Ringrose, L ;
Angrand, PO ;
Stewart, AF .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1997, 248 (03) :903-912
[15]   High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP [J].
Rodríguez, CI ;
Buchholz, F ;
Galloway, J ;
Sequerra, R ;
Kasper, J ;
Ayala, R ;
Stewart, AF ;
Dymecki, SM .
NATURE GENETICS, 2000, 25 (02) :139-140
[16]   DNA recombination with a heterospecific Cre homolog identified from comparison of the pac-c1 regions of P1-related phages [J].
Sauer, B ;
McDermott, J .
NUCLEIC ACIDS RESEARCH, 2004, 32 (20) :6086-6095
[17]   Efficient FLP recombination in mouse ES cells and oocytes [J].
Schaft, J ;
Ashery-Padan, R ;
van der Hoeven, F ;
Gruss, P ;
Stewart, AF .
GENESIS, 2001, 31 (01) :6-10
[18]   A reliable lacZ expression reporter cassette for multipurpose, knockout-first alleles [J].
Testa, G ;
Schaft, J ;
van der Hoeven, F ;
Glaser, S ;
Anastassiadis, K ;
Zhang, YM ;
Hermann, T ;
Stremmel, W ;
Stewart, AF .
GENESIS, 2004, 38 (03) :151-158
[19]   High-Efficient FLPo Deleter Mice in C57BL/6J Background [J].
Wu, Yingjie ;
Wang, Chunxin ;
Sun, Hui ;
LeRoith, Derek ;
Yakar, Shoshana .
PLOS ONE, 2009, 4 (11)
[20]   A new logic for DNA engineering using recombination in Escherichia coli [J].
Zhang, YM ;
Buchholz, F ;
Muyrers, JPP ;
Stewart, AF .
NATURE GENETICS, 1998, 20 (02) :123-128