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| Establishment of CRISPR/Cas9-edited FGF5 Cell Strains in Cashmere Goat |
| A Li ma, GAO Yuan, SU Xiao-hu, ZHOU Huan-min |
| College of Life Science, Inner Mongolia Agricultural University, Key Laboratory of Bio-Manufacturing of Inner Mongolia Autonomous Region, Hohhot 010018, China |
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Abstract To establish CRISPR/Cas9-edited FGF5 cell strains, we designed gRNA targeted sites around the first extra of the FGF5 gene and constructed two vectors by Cas/gRNA plasmid construction kit weredesigned. vectors into cashmere goat fibroblasts by electroporation respectively was transfected. T7 endonuclease 1 (T7E1) was used for the detection of mutation efficiency. The best vector was transfected into cashmere goat fetal fibroblasts and all the monoclones was cultured. Then all the cell colonies by sequencing were idenfified. Sequencing results demonstrated that CRISPR/Cas9 was available for FGF5 gene edited and 20 FGF5+/- and FGF5-/- cell colonies were obtained, and the effiency was 14.81%. The double mutated cell colonies could be used as donor cells to construct reconstructed embryos which provide the possibility in production of FGF5 edited cashmere goat in the future.
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Received: 07 January 2016
Published: 02 March 2016
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