Germ cell specific promoter drives ectopic transgene expression during embryogenesis

被引:12
作者
Bhullar, B
Schimidt, JV
Truong, T
Rancourt, D
Van der Hoorn, FA
机构
[1] Univ Calgary, Dept Biochem & Mol Biol, Calgary, AB T2N 4N1, Canada
[2] Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA
基金
加拿大健康研究院;
关键词
Cre recombinase; transgenic mice; Pgk2; promoter; spermatogenesis;
D O I
10.1002/mrd.1003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
In this study, we used the male germ cell-specific phosphoglycerate kinase 2 (Pgk2) promoter to generate Pgk2Cre transgenic mice to allow investigation of genes critically involved in meiosis. The Pgk2 promoter had been used previously to target transgene expression to spermatocytes and spermatids in several laboratories including ours. In several Cre targeting experiments using other promoters, ectopic Cre expression had been observed, but the timing and extent of this expression was not analyzed. We demonstrate that in adult mice the Pgk2Cre transgene is expressed specifically in spermatocytes and spermatids, as expected. However, in offspring from matings of Pgk2Cre mice and an H19loxP indicator strain, we discovered that recombination events had occurred in several, but not all, tissues to varying extents. The lacZ-loxP transgenic indicator strain was next used to uncover ectopic Cre expression even in single cells, which indicated that the Pgk2Cre transgene is expressed between days 11 and 15 during embryogenesis in several tissues and organs. Using an RT PCR assay we were unable to detect endogenous Pgk2 mRNA during embryogenesis or in adult tissues other than testis. In conclusion, the Pgk2 promoter is a valid choice for targeting gene expression to meiotic male germ cells, since transient ectopic expression is unlikely to have a discernable effect in most studies, but it may be inappropriate for utilization with Cre recombinase. Mol. Reprod. Dev. 59:25-32, 2001. (C) 2001 Wiley-Liss, Inc.
引用
收藏
页码:25 / 32
页数:8
相关论文
共 32 条
[1]
CLONING AND EXPRESSION OF THE MOUSE PGK-1 GENE AND THE NUCLEOTIDE-SEQUENCE OF ITS PROMOTER [J].
ADRA, CN ;
BOER, PH ;
MCBURNEY, MW .
GENE, 1987, 60 (01) :65-74
[2]
Cre-mediated somatic site-specific recombination in mice [J].
Akagi, K ;
Sandig, V ;
Vooijs, M ;
VanderValk, M ;
Giovannini, M ;
Strauss, M ;
Berns, A .
NUCLEIC ACIDS RESEARCH, 1997, 25 (09) :1766-1773
[3]
Spermatocyte-specific gene excision by targeted expression of Cre recombinase [J].
Ando, H ;
Haruna, Y ;
Miyazaki, J ;
Okabe, M ;
Nakanishi, Y .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2000, 272 (01) :125-128
[4]
HEART AND BONE-TUMORS IN TRANSGENIC MICE [J].
BEHRINGER, RR ;
PESCHON, JJ ;
MESSING, A ;
GARTSIDE, CL ;
HAUSCHKA, SD ;
PALMITER, RD ;
BRINSTER, RL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (08) :2648-2652
[5]
Transgenic Cre recombinase expression in germ cells and early embryogenesis directs homogeneous and ubiquitous deletion of loxP-flanked gene segments [J].
Bergqvist, I ;
Eriksson, B ;
Eriksson, M ;
Holmberg, D .
FEBS LETTERS, 1998, 438 (1-2) :76-80
[6]
Bypass of lethality with mosaic mice generated by Cre-loxP-mediated recombination [J].
Betz, UAK ;
Vosshenrich, CAJ ;
Rajewsky, K ;
Muller, W .
CURRENT BIOLOGY, 1996, 6 (10) :1307-1316
[7]
THE TESTIS-SPECIFIC PHOSPHOGLYCERATE KINASE GENE PGK-2 IS A RECRUITED RETROPOSON [J].
BOER, PH ;
ADRA, CN ;
LAU, YF ;
MCBURNEY, MW .
MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (09) :3107-3112
[8]
Targeting genes for self-excision in the germ line [J].
Bunting, M ;
Bernstein, KE ;
Greer, JM ;
Capecchi, MR ;
Thomas, KR .
GENES & DEVELOPMENT, 1999, 13 (12) :1524-1528
[9]
PROTEIN-DNA INTERACTIONS ASSOCIATED WITH THE ONSET OF TESTIS-SPECIFIC EXPRESSION OF THE MAMMALIAN PGK-2 GENE [J].
GEBARA, MM ;
MCCARREY, JR .
MOLECULAR AND CELLULAR BIOLOGY, 1992, 12 (04) :1422-1431
[10]
A FACTOR STIMULATING TRANSCRIPTION OF THE TESTIS-SPECIFIC PGK-2 GENE RECOGNIZES A SEQUENCE SIMILAR TO THE BINDING-SITE FOR A TRANSCRIPTION INHIBITOR OF THE SOMATIC-TYPE PGK-1 GENE [J].
GOTO, M ;
MASAMUNE, Y ;
NAKANISHI, Y .
NUCLEIC ACIDS RESEARCH, 1993, 21 (02) :209-214