Manipulating the mouse genome to engineer precise functional syntenic replacements with human sequence.
We have devised a strategy (called recombinase-mediated genomic replacement, RMGR) to allow the replacement of large segments (>100 kb) of the mouse genome with the equivalent human syntenic region. The technique involves modifying a mouse ES cell chromosome and a human BAC by inserting heterotypic lox sites to flank the proposed exchange interval and then using Cre recombinase to achieve segmental exchange. We have demonstrated the feasibility of this approach by replacing the mouse alpha globin regulatory domain with the human syntenic region and generating homozygous mice that produce only human alpha globin chains. Furthermore, modified ES cells can be used iteratively for functional studies, and here, as an example, we have used RMGR to produce an accurate mouse model of human alpha thalassemia. RMGR has general applicability and will overcome limitations inherent in current transgenic technology when studying the expression of human genes and modeling human genetic diseases.
Wallace
Helen A C
HA
Institute for Stem Cell Research, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JQ, United Kingdom.
Marques-Kranc
Fatima
F
Richardson
Melville
M
Luna-Crespo
Francisco
F
Sharpe
Jackie A
JA
Hughes
Jim
J
Wood
William G
WG
Higgs
Douglas R
DR
Smith
Andrew J H
AJ
D000818:Animals; D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D033481:Chromosomes, Mammalian / Q000235:genetics; D018390:Gene Targeting; D005818:Genetic Engineering; D016678:Genome / Q000235:genetics*; D005914:Globins / Q000235:genetics; D006403:Hematologic Tests; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008822:Mice, Transgenic; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017384:Sequence Deletion; D026801:Synteny / Q000235:genetics*
D005914:Globins