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Manipulating the mouse genome to engineer precise functional syntenic replacements with human sequence.

TitleManipulating the mouse genome to engineer precise functional syntenic replacements with human sequence.
Publication TypeJournal Article
Year of Publication2007
AuthorsWallace HAC, Marques-Kranc F, Richardson M, Luna-Crespo F, Sharpe JA, Hughes J, Wood WG, Higgs DR, Smith AG
JournalCell
Volume128
Issue1
Pagination197-209
Date Published2007 Jan 12
ISSN0092-8674
KeywordsAnimals, Base Sequence, Chromosomes, Artificial, Bacterial, Chromosomes, Mammalian, Gene Targeting, Genetic Engineering, Genome, Globins, Hematologic Tests, Humans, In Situ Hybridization, Fluorescence, Mice, Mice, Transgenic, Regulatory Sequences, Nucleic Acid, Sequence Deletion, Synteny
Abstract

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.

DOI10.1016/j.cell.2006.11.044
Alternate JournalCell
PubMed ID17218265
Grant ListMC_U137961144 / / Medical Research Council / United Kingdom
MC_U137961145 / / Medical Research Council / United Kingdom
Publication institute
CRM