Production of green fluorescent protein transgenic embryonic stem cells using the GENSAT bacterial artificial chromosome library.

TitleProduction of green fluorescent protein transgenic embryonic stem cells using the GENSAT bacterial artificial chromosome library.
Publication TypeJournal Article
Year of Publication2007
AuthorsTomishima MJ, Hadjantonakis A-K, Gong S, Studer L
JournalStem Cells
Volume25
Issue1
Pagination39-45
Date Published2007 Jan
ISSN1066-5099
KeywordsAnimals, Cell Differentiation, Chromosomes, Artificial, Bacterial, Electroporation, Embryonic Stem Cells, Escherichia coli, Fibroblasts, Flow Cytometry, Genes, Reporter, Green Fluorescent Proteins, Mice, Mice, Transgenic, Neurons
Abstract

Transgenic green fluorescent protein (GFP) reporter embryonic stem (ES) cells are powerful tools for studying gene regulation and lineage choice during development. Here we present a rapid method for the generation of ES cells expressing GFP under the control of selected genes. Bacterial artificial chromosomes (BACs) from a previously constructed GFP transcriptional fusion library (Gene Expression Nervous System Atlas [GENSAT]) were modified for use in ES cells, and multiple BAC transgenic ES cell lines were generated. Specific GFP expression in transgenic cell lines was confirmed during neural differentiation marking neural stem cells, neuronal precursors, and glial progeny by Hes5, Dll1, and GFAP, respectively. GFP was dynamically regulated in ES cell progeny in response to soluble factors that inhibit Notch signaling and a factor that directs astroglial fate choice. Our protocols provide a simple and efficient strategy to utilize the whole GENSAT BAC library to create hundreds of novel fluorescent cell lines for use in ES cell biology.

DOI10.1634/stemcells.2006-0173
Alternate JournalStem Cells
PubMed ID16990587
PubMed Central IDPMC2881625
Grant ListP30 CA008748 / CA / NCI NIH HHS / United States
R01 HD052115 / HD / NICHD NIH HHS / United States
R01 HD052115-03 / HD / NICHD NIH HHS / United States