Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2003-11-12
pubmed:abstractText
Previously, we designed a chromosomal vector (CV) and reported germline transmission of the vector by mice and regulated expression of the human tissue factor (F3) gene present on the CV. Further characterization and development of the CV are presented here. Mice could be bred with one to four copies of the CV per cell, and it is shown that F3 expression is proportional to the CV copy number. The insertion of large sequences into the CV was investigated by the insertion of a PAC, carrying 62.5 kb of human genomic DNA containing the CSN2 and STATH genes, into the CV by means of Cre/loxP recombination (CV(PAC)). Retrofitting the PAC with a cytomegalovirus (CMV)-5'HPRT/loxP cassette in Escherichia coli allowed efficient selection of CVs with PAC insert. Mitotic loss rates of the CV(PAC) were similar to the original CV. Furthermore, germline transmission efficiency and mitotic stability of the CV(PAC) in mice were not compromised. The human CSN2 and STATH genes were not expressed in the transchromosomal mice. In contrast, F3, already present on the CV, was expressed in CV(PAC)(+) F(1) mice similar to in CV(+) mice, suggesting that the insertion of large sequences does not interfere with transcription of genes present on the CV.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0888-7543
pubmed:author
pubmed:issnType
Print
pubmed:volume
82
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
596-605
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:14611801-Animals, pubmed-meshheading:14611801-Blotting, Western, pubmed-meshheading:14611801-Brain, pubmed-meshheading:14611801-Cells, Cultured, pubmed-meshheading:14611801-Chromosomes, Artificial, P1 Bacteriophage, pubmed-meshheading:14611801-Cytomegalovirus, pubmed-meshheading:14611801-DNA Primers, pubmed-meshheading:14611801-Electrophoresis, Gel, Pulsed-Field, pubmed-meshheading:14611801-Escherichia coli, pubmed-meshheading:14611801-Gene Dosage, pubmed-meshheading:14611801-Gene Expression Regulation, pubmed-meshheading:14611801-Gene Transfer Techniques, pubmed-meshheading:14611801-Genetic Vectors, pubmed-meshheading:14611801-In Situ Hybridization, Fluorescence, pubmed-meshheading:14611801-Meiosis, pubmed-meshheading:14611801-Mice, pubmed-meshheading:14611801-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:14611801-Thromboplastin, pubmed-meshheading:14611801-Transgenes
pubmed:year
2003
pubmed:articleTitle
Controlled transgene dosage and PAC-mediated transgenesis in mice using a chromosomal vector.
pubmed:affiliation
Department of Human Genetics, Flanders Interuniversity Institute for Biotechnology, Herestraat 49, University of Leuven, Louvain B-3000, Belgium.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't