Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2004-1-27
pubmed:abstractText
We have developed a gene trap vector that transduces an EGFP-neo fusion gene (Eno) to monitor the expression of trapped genes in living cells and embryos. Upon in vitro differentiation, most gene-trapped embryonic stem (ES) cell clones exhibited detectable green fluorescence in various specialized cell types, which can be followed in the live culture in real time. Populations of ES cell-derived cardiomyocytes, smooth muscle cells, vascular endothelial cells, and hematopoietic cells were readily recognized by their distinctive morphologies coupled with unique activities, allowing efficient screening for clones with trapped genes expressed in cardiovascular lineages. Applying G418 selection in parallel differentiation cultures further increased detection sensitivity and screening throughput by enriching reporter-expressing cells with intensified green fluorescent protein signals. Sequence analyses and chimera studies demonstrated that the expression of trapped genes in vivo closely correlated with the observed lineage specificity in vitro. This provides a strategy to identify and mutate genes expressed in lineages of interest for further functional studies.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
1058-8388
pubmed:author
pubmed:copyrightInfo
Copyright 2004 Wiley-Liss, Inc.
pubmed:issnType
Print
pubmed:volume
229
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
319-27
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:14745956-Animals, pubmed-meshheading:14745956-Artificial Gene Fusion, pubmed-meshheading:14745956-Cardiovascular System, pubmed-meshheading:14745956-Cell Culture Techniques, pubmed-meshheading:14745956-Cell Differentiation, pubmed-meshheading:14745956-Cell Lineage, pubmed-meshheading:14745956-Endothelium, Vascular, pubmed-meshheading:14745956-Female, pubmed-meshheading:14745956-Gene Expression Profiling, pubmed-meshheading:14745956-Gene Expression Regulation, Developmental, pubmed-meshheading:14745956-Green Fluorescent Proteins, pubmed-meshheading:14745956-Luminescent Proteins, pubmed-meshheading:14745956-Male, pubmed-meshheading:14745956-Mice, pubmed-meshheading:14745956-Myocytes, Cardiac, pubmed-meshheading:14745956-Myocytes, Smooth Muscle, pubmed-meshheading:14745956-Recombinant Proteins, pubmed-meshheading:14745956-Stem Cells, pubmed-meshheading:14745956-Transfection, pubmed-meshheading:14745956-Transplantation Chimera
pubmed:year
2004
pubmed:articleTitle
Differentiation trapping screen in live culture for genes expressed in cardiovascular lineages.
pubmed:affiliation
Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan, USA. chenz@zgi.com
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't