Coding sequences for a cerulean (CFP) fluorescent protein gene were inserted into the exchange cassette.  In this manner, CFP is constituitely expressed under control of the endogenous ROSA26 promoter.  The exchange plasmid also contains a 51bp translational enhancer (5' leader sequence from Xenopus beta-globin), a Kozak sequence upstream of the start codon, and intronic a polyA sequences from the rabbit beta-globin gene that confer stability to the mRNA.

 


Record History
Added on December 21, 2011 at 3:59 PM by Peters, Jody
Modified on April 13, 2017 at 3:07 PM by Skelton, Jennifer
Shared with (contributions)
Public: Shared Collection

Mouse Information

Common Name Rosa26CFPMgn
Research Applications Fluorescent proteins
MMRRC ID 036288-MU
Jackson Laboratories Stock No Not provided
VUCMR ID NH
Additional Strain Information Not provided

Genetic Alteration

Mutation #1: RMCE Targeted Mutagenesis
Type of AlleleCassette Acceptor
Targeted GeneName: gene trap ROSA26, Philippe Soriano
Symbol: Gt(ROSA)26Sor
NCBI: 14910
Allele Name: targeted mutation 1
Symbol: Rosa26
MGI: 5287869
Description of Targeting Vector

This mouse line contains a loxed cassette acceptor (LCA) allele in which a 5.17 kb region of the gene has been replaced by a lox71 site, a puromycin-(delta)-thymidine kinase fusion gene driven by the mouse phosphoglycerol kinase promoter, a kanamycin resistance gene driven by the bacterial EM7 promoter, and a lox2272 site.  These features enable use of Recombinase-Mediated Cassette Exchange (RMCE) for the rapid insertion of various DNAs into the Rosa26 gene locus. 


Vector Genbank File pRosa26.LCA.gb
Allele Map Not Provided
PCR Genotyping Protocol Not provided
Type of Allele Global Null
Exchanged Cassette Gene Name ()
Exchanged Cassette Allele Name pRosa.EN.CFP.bGsplicepA.Neo
Description of Exchange Vector

The pRosa.EN.CFP.bGsplicepA.Neo exchange vector was made on a backbone of a basal exchange vector which contains a 4.081 kb sequence from the Rosa26 locus, Lox71/Lox2272 sites, and a flrted (flanked by FRT) Pgk-Neo cassette that is used for positive selection of ES cells after RMCE.  A 5' translational enhancer-CFP-intron-containing rabbit beta-globin polyA sequence was inserted (in place of Rosa26 exon 1), between the Lox71 and the Pgk-Neo sites.

Exchanged Cassette Genbank File pRosa.EN.CFP.bGsplicepA.neo1.gb
Citations
Publication
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 11/8/2013)
PMID: 21324933

Background Strain Information

Strain Type Mixed
Chimera/Founder Genetic Background 129S6/SvEvTac
Current Genetic Background C57BL/6J
Number of Generations Backcrossed 4
Strain Description

Founder chimeras were crossed with C57Bl/6J for 4 generations.

Attachments

Document 6_colors_of_mice.jpg - Added on October 23, 2013 at 4:03 PM by Anna Osipovich

The images of newborn offspring from intercrosses of Rosa26Cherry/ ,Rosa26CFP/ and Rosa26YFP/ mice were taken with a stereoscope using CFP, YFP and RFP filter and subsequently overlayed.


Document CFPalone2.jpg - Added on October 23, 2013 at 4:02 PM by Anna Osipovich

The image of Rosa26CFP/ mouse was taken with a stereoscope using CFP filter.


Document R26_EN_CFP_RMCEs.png - Added on February 7, 2012 at 1:36 PM by Jody Peters

Linear Map of Targeting Event