Selection of targeted mutants from a library of randomly mutagenized ES cells
- PMID: 21080287
- DOI: 10.1007/978-1-60761-974-1_17
Selection of targeted mutants from a library of randomly mutagenized ES cells
Abstract
A method for random relatively unbiased mutagenesis of ES cells with a mutagenic retroviral vector is described. An orderly assembly of mutant ES cells in multi-well plates is generated. 3D pooling of the wells of the assembly allows quick PCR search for insertions in genes of interest. Mutant ES cell clones are then isolated from the positive wells and used to produce mutant animals using conventional techniques.
Similar articles
-
The loss-of-allele assay for ES cell screening and mouse genotyping.Methods Enzymol. 2010;476:295-307. doi: 10.1016/S0076-6879(10)76017-1. Methods Enzymol. 2010. PMID: 20691873
-
Targeted gene modification in mouse ES cells using integrase-defective lentiviral vectors.Genesis. 2009 Apr;47(4):217-23. doi: 10.1002/dvg.20469. Genesis. 2009. PMID: 19208434
-
Construction of engineered murine embryonic stem cells with conditional knockout of FGFR2 depending on Cre-loxP.Biocell. 2006 Aug;30(2):269-78. Biocell. 2006. PMID: 16972551
-
Gene-trap mutagenesis: past, present and beyond.Nat Rev Genet. 2001 Oct;2(10):756-68. doi: 10.1038/35093548. Nat Rev Genet. 2001. PMID: 11584292 Review.
-
Animal transgenesis: an overview.Brain Struct Funct. 2010 Mar;214(2-3):91-109. doi: 10.1007/s00429-009-0230-8. Epub 2009 Nov 25. Brain Struct Funct. 2010. PMID: 19937345 Review.
MeSH terms
LinkOut - more resources
Full Text Sources