Linked Life Data

10471751

Source:http://linkedlifedata.com/resource/pubmed/id/10471751

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
18
pubmed:dateCreated
1999-9-15
pubmed:abstractText
We have applied Cre-mediated double lox recombination to embryonic stem (ES) cells to facilitate repeated knock-ins at a target locus, thus helping to assure correct temporal and spatial transgene expression in mice. Using homologous recombination, we inserted a double lox cassette a few nucleo-tides before the authentic ATG start of MHL-1, the gene coding for the major subunit of the asialoglycoprotein receptor. The cassette carries a marker gene bounded by heterospecific lox sites that cannot recombine with each other, but which can undergo recombination with like sites on an incoming double lox targeting vector. Cre-mediated replacement of the lox-delimited genomic segment at MHL-1 with targeting DNA occurs at a frequency three times that of random DNA integration and is sufficiently robust that correctly targeted ES colonies can be identified by PCR screening without relying on any drug selection procedure. Moreover, double lox recombination at the MHL-1 locus is not dependent on the presence or absence of a transcriptionally active promoter at the genomic target. The strategy and vectors described here are generally applicable to designing double lox targeted knock-ins at any locus in ES cells and should prove useful in more precise molecular engineering of the mouse genome.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
1362-4962
pubmed:author
pubmed:issnType
Electronic
pubmed:day
15
pubmed:volume
27
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
e21
pubmed:dateRevised
2008-11-20
pubmed:meshHeading
pubmed:year
1999
pubmed:articleTitle
Segmental genomic replacement in embryonic stem cells by double lox targeting.
pubmed:affiliation
Laboratory of Biochemistry and Metabolism, National Institute of Diabetes, Digestive and Kidney Disease, 9000 Rockville Pike, Bethesda, MD 20892, USA.
pubmed:publicationType
Journal Article