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Direct removal in the mouse of a floxed neo gene from a three‐loxp conditional knockout allele by two novel approaches

Direct removal in the mouse of a floxed neo gene from a three‐loxp conditional knockout allele by... Summary: The presence in an intron of the ploxP‐neo‐loxP cassette often results in severe interference with gene expression. Consequently, many investigators selectively remove the ploxP‐neo‐loxP cassette by transient expression of Cre in ES cells. Although effective, the added manipulation of the ES cells may reduce the likelihood that a clone will be able to transmit via the germline. Therefore, we developed two novel approaches that remove the ploxP‐neo‐loxP by Cre‐mediated recombination in mouse. First, the ploxP‐neo‐loxP‐containing mice were crossed with EIIa‐Cre transgenic mice. Second, a Cre‐expression plasmid was injected into pronuclei of fertilized eggs bearing the ploxP‐neo‐loxP allele. Both approaches produced mosaic mice with partial and complete excision. These mosaic mice were then mated, and the neo‐less conditional knockout allele was found in the offspring after screening only a few litters. These procedures provide options for removing neo directly in the mouse in addition to the commonly used approach that deletes neo in ES cells. genesis 30:1–6, 2001. © 2001 Wiley‐Liss, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Genesis: the Journal of Genetics and Development Wiley

Direct removal in the mouse of a floxed neo gene from a three‐loxp conditional knockout allele by two novel approaches

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References (21)

Publisher
Wiley
Copyright
Copyright © 2001 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1526-954X
eISSN
1526-968X
DOI
10.1002/gene.1025
Publisher site
See Article on Publisher Site

Abstract

Summary: The presence in an intron of the ploxP‐neo‐loxP cassette often results in severe interference with gene expression. Consequently, many investigators selectively remove the ploxP‐neo‐loxP cassette by transient expression of Cre in ES cells. Although effective, the added manipulation of the ES cells may reduce the likelihood that a clone will be able to transmit via the germline. Therefore, we developed two novel approaches that remove the ploxP‐neo‐loxP by Cre‐mediated recombination in mouse. First, the ploxP‐neo‐loxP‐containing mice were crossed with EIIa‐Cre transgenic mice. Second, a Cre‐expression plasmid was injected into pronuclei of fertilized eggs bearing the ploxP‐neo‐loxP allele. Both approaches produced mosaic mice with partial and complete excision. These mosaic mice were then mated, and the neo‐less conditional knockout allele was found in the offspring after screening only a few litters. These procedures provide options for removing neo directly in the mouse in addition to the commonly used approach that deletes neo in ES cells. genesis 30:1–6, 2001. © 2001 Wiley‐Liss, Inc.

Journal

Genesis: the Journal of Genetics and DevelopmentWiley

Published: May 1, 2001

Keywords: ; ; ; ;

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