Metaphase FISHing of transgenic mice recommended: FISH and SKY define BAC-mediated balanced translocation

被引:14
作者
Abrahams, BS
Chong, ACO
Nisha, M
Milette, D
Brewster, DA
Berry, ML
Muratkhodjaev, F
Mai, S
Rajcan-Separovic, E
Simpson, EM
机构
[1] Univ British Columbia, Dept Med Genet, Vancouver, BC V5Z 4H4, Canada
[2] British Columbia Res Inst Childrens & Womens Hlth, Ctr Mol Med & Therapeut, Vancouver, BC, Canada
[3] Univ British Columbia, Ctr Childrens & Womens Hlth, Dept Pathol Cytogenet, Vancouver, BC, Canada
[4] Univ Manitoba, Manitoba Inst Cell Biol, CancerCare Manitoba, Genom Ctr Canc Res & Diag, Winnipeg, MB, Canada
[5] Jackson Lab, Bar Harbor, ME 04609 USA
关键词
balanced translocation; bacterial artificial chromosome; transgenic mice; breakpoints; Nr2e1; FISH; SKY; tailless; orphan receptor;
D O I
10.1002/gene.10205
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The evolving trend to use larger transgenes and their associated increased chance of unexpected genetic events mandates more careful characterization of transgenic mice. In characterizing our five new mouse strains transgenic for the BAC, bEMS4, we have identified the highest copy number reported to date: the stable incorporation of similar to40 copies of a 194-kb expressed transgene in a single insertion site. We caution, however, that standard molecular techniques failed to identify a balanced translocation in another strain, and an inappropriate site of insertion in a third. Molecular cytogenetic analysis using metaphase FISH was the minimum level of characterization needed to reveal these unexpected genetic events. In addition, we combined FISH and SKY to identify the transgene at the breakpoints of the balanced translocation, t(3;9). This is the first description of a BAC-mediated chromosomal rearrangement and the first application of SKY to identify transgene-induced chromosomal rearrangements. genesis 36:134-141, 2003. (C) 2003 Wiley-Liss, Inc.
引用
收藏
页码:134 / 141
页数:8
相关论文
共 29 条
[21]   FISH probes for mouse chromosome identification [J].
Shi, YP ;
Mohapatra, G ;
Miller, J ;
Hanahan, D ;
Lander, E ;
Gold, P ;
Pinkel, D ;
Gray, J .
GENOMICS, 1997, 45 (01) :42-47
[22]   AN INTERSTITIAL DELETION IN MOUSE-Y CHROMOSOMAL DNA CREATED A TRANSCRIBED ZFY FUSION GENE [J].
SIMPSON, EM ;
PAGE, DC .
GENOMICS, 1991, 11 (03) :601-608
[23]   Physical characterization of the chromosomal rearrangements that accompany the transgene insertion in the chakragati mouse mutant [J].
Smiraglia, DJ ;
Ratty, AK ;
Gross, KW .
GENOMICS, 1997, 45 (03) :562-571
[24]   PRENATAL LETHALITIES IN MICE HOMOZYGOUS FOR HUMAN GROWTH-HORMONE GENE-SEQUENCES INTEGRATED IN THE GERM LINE [J].
WAGNER, EF ;
COVARRUBIAS, L ;
STEWART, TA ;
MINTZ, B .
CELL, 1983, 35 (03) :647-655
[25]   POSITION EFFECTS ON EUKARYOTIC GENE-EXPRESSION [J].
WILSON, C ;
BELLEN, HJ ;
GEHRING, WJ .
ANNUAL REVIEW OF CELL BIOLOGY, 1990, 6 :679-714
[26]   The combination of SKY and specific loci detection with FISH or immunostaining [J].
Ye, CJ ;
Lu, W ;
Liu, G ;
Bremer, SW ;
Wang, YA ;
Moens, P ;
Hughes, M ;
Krawetz, SA ;
Heng, HHQ .
CYTOGENETICS AND CELL GENETICS, 2001, 93 (3-4) :195-202
[27]   Fierce:: a new mouse deletion of Nr2e1;: violent behaviour and ocular abnormalities are background-dependent [J].
Young, KA ;
Berry, ML ;
Mahaffey, CL ;
Saionz, JR ;
Hawes, NL ;
Chang, B ;
Zheng, QY ;
Smith, RS ;
Bronson, RT ;
Nelson, RJ ;
Simpson, EM .
BEHAVIOURAL BRAIN RESEARCH, 2002, 132 (02) :145-158
[28]   Unrepaired DNA breaks in p53-deficient cells lead to oncogenic gene amplification subsequent to translocations [J].
Zhu, CM ;
Mills, KD ;
Ferguson, DO ;
Lee, C ;
Manins, J ;
Fleming, J ;
Gao, YJ ;
Morton, CC ;
Alt, FW .
CELL, 2002, 109 (07) :811-821
[29]   REGIONAL LOCALIZATION OF THE FIBRONECTIN AND GAMMA-CRYSTALLIN GENES TO MOUSE CHROMOSOME-1 BY INSITU HYBRIDIZATION [J].
ZNEIMER, SM ;
WOMACK, JE .
CYTOGENETICS AND CELL GENETICS, 1988, 48 (04) :238-241