NDLI: Construction of Gene-Targeting Vectors: a Rapid Mu in vitro DNA Transposition-Based Strategy Generating Null, Potentially Hypomorphic, and Conditional Alleles

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Construction of Gene-Targeting Vectors: a Rapid Mu in vitro DNA Transposition-Based Strategy Generating Null, Potentially Hypomorphic, and Conditional Alleles

Acknowledgement ( Transgenic Research , Volume 10 , Issue 1 )

Transgenic Research : Volume 9

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Transgenic Research : Volume 6

Construction of Gene-Targeting Vectors: a Rapid Mu in vitro DNA Transposition-Based Strategy Generating Null, Potentially Hypomorphic, and Conditional Alleles

Content Provider	Springer Nature Link
Author	Vilen, Heikki Eerikäinen, Simo Tornberg, Janne Airaksinen, Matti S. Savilahti, Harri
Copyright Year	2001
Abstract	Gene targeting into mammalian genomes by means of homologous recombination is a powerful technique for analyzing gene function through generation of transgenic animals. Hundreds of mouse strains carrying targeted alleles have already been created and recent modifications of the technology, in particular generation of conditional alleles, have extended the usefulness of the methodology for a variety of special purposes. Even though the standard protocols, including the construction of gene-targeting vector plasmids, are relatively straightforward, they typically involve time-consuming and laborious gene mapping and/or sequencing steps. To produce various types of gene-targeting constructions rapidly and with minimum effort, we developed a strategy, that utilizes a highly efficient in vitro transposition reaction of phage Mu, and tested it in a targeting of the mouse Kcc2 gene locus. A vast number and different types of targeting constructions can be generated simultaneously with little or no prior sequence knowledge of the gene locus of interest. This quick and efficient general strategy will facilitate easy generation of null, potentially hypomorphic, and conditional alleles. Especially useful it will be in the cases when effects of several exons within a given gene are to be studied, a task that necessarily will involve generation of multiple constructions. The strategy extends the use of diverse recombination reactions for advanced genome engineering and complements existing recombination-based approaches for generation of gene-targeting constructions.
Starting Page	69
Ending Page	80
Page Count	12
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	10
Issue Number	1
e-ISSN	15739368
Language	English
Publisher	Kluwer Academic Publishers-Plenum Publishers
Publisher Date	2001-01-01
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Human Genetics Plant Sciences Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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