An RMCE-derived cyan fluorescent protein reporter allele for Pdx1

• Published in: Genesis (New York, N.Y. : 2000) Vol. 50; no. 4
• Main Authors: Potter, Leah A., Choi, Eunyoung, Hipkens, Susan B., Wright, Christopher V.E., Magnuson, Mark A.
• Format: Journal Article
• Language: English
• Published: 06-01-2012
• ISSN: 1526-954X; 1526-968X
• DOI: 10.1002/dvg.20804

Fluorescent protein (FP) reporter alleles are useful both for identifying and purifying specific cell populations in the mouse. Here, we report the generation of mouse embryonic stem cells that contain a
Pdx1
loxed cassette acceptor (
Pdx1
LCA
) allele and the use of recombinase-mediated cassette exchange (RMCE) to derive mice that contain a
Pdx1
CFP
(Cerulean) reporter allele. Mice with this allele exhibited cyan fluorescence within the previously well-characterized
Pdx1
expression domain in posterior foregut endoderm. Immunolabeling showed that endogenous Pdx1 was co-expressed with CFP at all time points examined. Furthermore, fluorescence-activated cell sorting (FACS) was used to isolate CFP-positive cells from E11.5 and E18.5 embryonic tissues using both 405 and 445 nm lasers, although the latter resulted in a nearly 50-fold increase in emission intensity. The
Pdx1
CFP
allele will enable the isolation of specific foregut endoderm and pancreatic cell populations, both alone and in combination with other FP reporter alleles.