- Function research of genes
- Function research of smooth muscle cell
Heterozygous mice for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. In this strain, iCre recombinase expression is under the control of Tagln promoter. When crossed with a strain containing a loxP-site flanked sequence of interest, Cre-mediated recombination will result in deletion of the floxed sequence in the offspring.
Gene editing strategy
ICRE, WPRE and PolyA cassette were inserted into the first intron of mouse endogenous transgelin gene to express iCre recombinase regulated by the Tagln promoter of mouse endogenous gene, and the PolyA element knocked out the Tagln gene in mice.The correct genotype mice were obtained by microinjection of mouse ES cells. This design is based on transcript-201 (ENSMUST00000034590.3, NM_011526 ) This gene encodes a smooth muscle cell-specific cytoskeletal protein. The encoded protein is structurally similar to calponin, an actin-binding protein. In mouse models of atherosclerosis the gene product may be involved in plaque cell and atherosclerotic lesion formation during atherogenesis.
The frozen sections of the stomach (a), aorta (b), jejunum (c) and bladder (d) of mice were detected by IF, respectively, and the results showed that tdTomato specific expression in Tagln-iCre (Mut /+);CAG-tdTomato (Mut / +) genotypes of double positive mice in alpha SMA markers of smooth muscle cells (with SMA protein markers of smooth muscle cells, the vast majority of the proportion of cells containing tdTomato red fluorescent protein signals, but it is not 100%, analyze the possible reasons may be the efficiency of the recombinant enzyme wasn’t 100%), can attest, iCre recombinant enzyme specificity expressed in smooth muscle cells, other tissue cells tdTomato red fluorescent protein signal wasn’t detected. The mice in the negative control group also did not express tdTomato red fluorescent protein.