Generation of PECAM-1 (CD31) conditional knockout mice. Genesis 2020 Feb;58(2):e23346
Date
11/16/2019Pubmed ID
31729819Pubmed Central ID
PMC7021573DOI
10.1002/dvg.23346Scopus ID
2-s2.0-85075415188 (requires institutional sign-in at Scopus site) 7 CitationsAbstract
Platelet endothelial cell adhesion molecule 1 (PECAM-1) is an adhesion and signaling receptor that is expressed on endothelial and hematopoietic cells and plays important roles in angiogenesis, vascular permeability, and regulation of cellular responsiveness. To better understanding the tissue specificity of PECAM-1 functions, we generated mice in which PECAM1, the gene encoding PECAM-1, could be conditionally knocked out. A targeting construct was created that contains loxP sites flanking PECAM1 exons 1 and 2 and a neomycin resistance gene flanked by flippase recognition target (FRT) sites that was positioned upstream of the 3' loxP site. The targeting construct was electroporated into C57BL/6 embryonic stem (ES) cells, and correctly targeted ES cells were injected into C57BL/6 blastocysts, which were implanted into pseudo-pregnant females. Resulting chimeric animals were bred with transgenic mice expressing Flippase 1 (FLP1) to remove the FRT-flanked neomycin resistance gene and mice heterozygous for the floxed PECAM1 allele were bred with each other to obtain homozygous PECAM1 flox/flox offspring, which expressed PECAM-1 at normal levels and had no overt phenotype. PECAM1 flox/flox mice were bred with mice expressing Cre recombinase under the control of the SRY-box containing gene 2 (Sox2Cre) promoter to delete the floxed PECAM1 allele in offspring (Sox2Cre;PECAM1 del/WT ), which were crossbred to generate Sox2Cre; PECAM1 del/del offspring. Sox2Cre; PECAM1 del/del mice recapitulated the phenotype of conventional global PECAM-1 knockout mice. PECAM1 flox/flox mice will be useful for studying distinct roles of PECAM-1 in tissue specific contexts and to gain insights into the roles that PECAM-1 plays in blood and vascular cell function.
Author List
Zhi H, Kanaji T, Fu G, Newman DK, Newman PJAuthors
Debra K. Newman PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinDebra K. Newman PhD Investigator in the Blood Research Institute department at BloodCenter of Wisconsin
MESH terms used to index this publication - Major topics in bold
AnimalsGene Knockout Techniques
Integrases
Mice
Mice, Inbred C57BL
Mouse Embryonic Stem Cells
Platelet Endothelial Cell Adhesion Molecule-1
SOXB1 Transcription Factors
