Abstract
Embryonic stem cell technology revolutionized biology by providing a means to assess mammalian gene function in vivo. Although it is now routine to generate mice from embryonic stem cells, one of the principal methods used to create mutations, gene targeting, is a cumbersome process. Here we describe the indexing of 93,960 ready-made insertional targeting vectors from two libraries. 5,925 of these vectors can be used directly to inactivate genes with an average targeting efficiency of 28%. Combinations of vectors from the two libraries can be used to disrupt both alleles of a gene or engineer larger genomic changes such as deletions, duplications, translocations or inversions. These indexed vectors constitute a public resource (Mutagenic Insertion and Chromosome Engineering Resource; MICER) for high-throughput, targeted manipulation of the mouse genome.
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Acknowledgements
We thank W.C. Skarnes, members of the laboratories of W.C. Skarnes and A.B. and the Sanger informatics team for discussions and assistance. D.J.A and L.v.d.W. were supported by a National Health and Medical Research Council CJ Martin fellowship and a National Health and Medical Research Council CJ Martin/RG Menizes fellowship, respectively. J.J. is a recipient of a NWO Genomics Fellowship from the Netherlands Organization for Scientific Research. This work was supported by the Wellcome Trust (UK).
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Supplementary information
Supplementary Table 1
Targeting frequency data for MICER vectors. (XLS 13 kb)
Supplementary Table 2
Sentinel genes list. (XLS 1040 kb)
Supplementary Table 3
MICER gene disruption alleles. (XLS 1328 kb)
Supplementary Table 4
Restriction enzymes that can be used to linearize MICER vectors. (PDF 3 kb)
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Adams, D., Biggs, P., Cox, T. et al. Mutagenic Insertion and Chromosome Engineering Resource (MICER). Nat Genet 36, 867–871 (2004). https://doi.org/10.1038/ng1388
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DOI: https://doi.org/10.1038/ng1388
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