Establishment Of Label-freed Talen-edited FGF5 Cell Strains In Cashmere Goat

Fibroblast growth factor 5 (FGF5) is one of the member of the FGFs gene family factor, it is important to the growth of the regulation of cell growth factor. The FGF5 has an important regulating role in hair growth cycle, FGF5 gene mutations can lead to the extension of hair growth cycle in the growth period, so as to increase the length of the hair.The purpose of this study was to establish a safe and effective method, design label-freed TALEN gene editing carrier, transfer label-freed carrier TALEN gene editing electricity method transfection into the fetal fibroblasts of cashmere goat, screening of positive cells and propagation, for somatic cell nuclear transfer without screening markers of donor cells, For the creation of a new germplasm materials of cashmere goat with high yield and velvet traits lay the foundation.According to the first exon FGF5 gene sequences of the cashmere goats, the researcher successfuliy design and build the three pairs of unmarked TALEN carrier pCS7-FGF5-eT1, pCS7-FGF5-eT2, pCS7-FGF5-eT3.Through sequencing,SSA activity detection and T7E1 enzyme identification, the results show that the vector was successfully constructed, and pCS7-FGF5-eT2 is high activity and available.With the electricity transfection method, have transfected active TALEN carrier pCS7-FGF5-eT2 into the fetal fibroblasts of the cashmere goats,choose positive cells with single cell culture method and expand breeding.After sequencing identification,2-29 single cell strains in the target area into the base,resulting in the target area downstream frameshift mutations of genes,which will result in FGFS gene can’t normal protein,constructed label-freed FGF5 gene transgenic cell strains of the cashmere goats,which laid a foundation for the production of label-freed transgenic cashmere goats.