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

   

Authors:	Tomishima, M. J.; Hadjantonakis, A. K.; Gong, S.; Studer, L.
Article Title:	Production of green fluorescent protein transgenic embryonic stem cells using the GENSAT bacterial artificial chromosome library
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. ©AlphaMed Press.
Keywords:	protein expression; flow cytometry; animals; mice; embryonic stem cell; green fluorescent protein; astrocyte; notch receptor; neural stem cell; cell differentiation; neurons; bacteria (microorganisms); bacterial artificial chromosome; chromosomes, artificial, bacterial; mice, transgenic; escherichia coli; progeny; transgene; fibroblasts; embryonic stem cells; green fluorescent proteins; electroporation; genes, reporter; nerve cell differentiation; fluorescent reporter; neural differentiation; bacterial artificial chromosome transgenesis
Journal Title:	Stem Cells
Volume:	25
Issue:	1
ISSN:	1066-5099
Publisher:	AlphaMed Press
Date Published:	2007-01-01
Start Page:	39
End Page:	45
Language:	English
DOI:	10.1634/stemcells.2006-0173
PUBMED:	16990587
PROVIDER:	scopus
PMCID:	PMC2881625
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-33846019592&partnerID=40&md5=703b526bcadcb4cdb0603c2a90585c43
Notes:	--- - "Cited By (since 1996): 14" - "Export Date: 17 November 2011" - "CODEN: STCEE" - "Source: Scopus"

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