Rosa26^EN-GFP-Neo - ES Cell Line RES2542

ESC Line Information

Cell Line Name:	Rosa26^EN-GFP-Neo
Parental Cell Line:	TL1 Rosa26{LCA} clone 5B9
Background Strain:	129
Culturing Protocol:	std_mesc_culture.doc
Description:	This ES cell line contains eGFP inserted into the Rosa26 gene locus. These cells were used to identify an optimal combination of regulatory elements for fluorescent protein expression from a single genomic copy and can serve as a reference cell line for fluorescent cell sorting.

Genetic Alterations

1) RMCE Targeted Mutagenesis

Type of Allele

Cassette Acceptor

Targeted Gene

gene trap ROSA 26, Philippe Soriano (Gt(ROSA)26Sor - NCBI GeneID:14910)

Targeted Allele

targeted mutation 1 (Rosa26^tm1(LCA) - MGI:104735)

Description of Targeting Vector

The Rosa 26 cassette acceptor allele was created by replacing a 5.165 kb region of this gene containing exon 1 with a floxed tk-neo cassette, a puromycin-delta-thymidine kinase fusion gene driven by the mouse phosphoglycerol kinase promoter (pU-deltaTK) and a neomycin resistant gene driven by the bacterial EM7 promoter (EM7neo) flanked by minimal (34 bp) tandemly oriented lox71 and lox2272 sites.

Targeting Vector Genbank File

pRosa26.LCA.gb

Recombinase-Mediated Cassette Exchange Stage

Type of Allele:

Gene Replacement

Exchanged Cassette Gene

green fluorescent protein (GFP)

Exchanged Cassette Allele Name

Rosa26^EN.GFP

Description of Exchange Vector

not available

Exchange Vector Genbank File:

prosa.en.gfp.bgsplicepa.neo.gb

Citations

PubMedID	Citation
21324933	Quantification of factors influencing fluorescent protein expression using RMCE to generate an allelic series in the ROSA26 locus in mice. (2011) Dis Model Mech 4: 537-47 (Added 2012-09-24 16:36:23.369844)

Associated Images

Image 1
	Description: A green fluorescent protein (eGFP) gene was placed under control of the endogenous Rosa 26 promoter. The exchange plasmid also has a 51 bp translational enhancer (5' leader sequence from Xenopus beta-globin gene), a Kozak sequence upstream of the start codon, and an intronic region and polyA site from the rabbit beta globin gene. Reference: Not provided

Repositories

Magnuson Lab
Request this resource	Stock #: Not provided Availability Notes: Not provided

Contact Information

Preferred Contact
Name	Mark Magnuson
Institution	Vanderbilt University
Phone	615-322-7006
Email	mark.magnuson@vanderbilt.edu
Primary Lab Contact
Name	Susan Hipkens
Institution	Vanderbilt University
Phone	615-343-7422
Email	susan.hipkens@vanderbilt.edu