%0 Journal Article %T General method for the modification of different BAC types and the rapid generation of BAC transgenic mice %+ Institute for Medical Microbiology, Immunology and Hygiene %+ Laboratory of Molecular Biology %+ Université de Lausanne = University of Lausanne (UNIL) %A Sparwasser, Tim %A Gong, Shiaoching %A Li, James Y.H. %A Eberl, Gerard %< avec comité de lecture %@ 1526-954X %J Genesis %I Wiley-Blackwell %V 38 %N 1 %P 39 - 50 %8 2004-01 %D 2004 %R 10.1002/gene.10249 %M 14755803 %K knockin %K recombination %K transgenic mice %K overlap PCR %K bacterial artificial chromosomes %Z Life Sciences [q-bio]/Immunology %Z Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology %Z Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN]Journal articles %X Most genome projects have relied on the sequencing of bacterial artificial chromosomes (BACs), which encompass 100-300 kb of genomic DNA. As a consequence, several thousand BAC clones are now mapped to the human and mouse genome. It is therefore possible to identify in silico a BAC clone that carries a particular gene and obtain it commercially. Given the large size of BACs, most if not all regulatory sequences of a gene are present and can be used to direct faithful and tissue-specific expression of heterologous genes in vitro in cell cultures and in vivo in BAC-transgenic mice. We describe here an optimized and comprehensive protocol to select, modify, and purify BACs in order to generate BAC-transgenic mice. Importantly, this protocol includes a method to generate, within 2 days, complex plasmid cassettes required to modify BACs, and to efficiently modify different types of BACs selected from the two major BAC libraries available. Altogether, using a combination of genomic database analysis, overlap PCR cloning, and BAC recombination in bacteria, our approach allows for the rapid and reliable generation of "pseudo knockin" mice. genesis 38:39-50, 2004. %G English %L pasteur-01402770 %U https://pasteur.hal.science/pasteur-01402770 %~ PASTEUR