Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6490817 | Journal of Biotechnology | 2015 | 19 Pages |
Abstract
Production of nuclear donor cells with a high percentage of desired modifications is a critical step in the successful generation of genetically modified pigs through somatic cell nuclear transfer (SCNT). The CRISPR/Cas9 system has been used for efficient modification of the nuclear DNA in eukaryotic cells, including porcine cells. However, in vitro modified cells are often phenotypically indistinguishable from unmodified cells, hampering their enrichment. Here we investigate a dual fluorescence selection system for the efficient enrichment of porcine embryonic fibroblasts (PEFs) with CRISPR/Cas9-induced chromosomal deletions. Enrichment of cells with 170Â bp deletions reached a frequency of 74%, whilst enrichment of cells with a larger 5Â kb deletions achieved a frequency of 46%. This demonstrates the utility of a dual fluorescence reporter as an attractive tool for improving the efficiency of generating genome edited pigs.
Keywords
Related Topics
Physical Sciences and Engineering
Chemical Engineering
Bioengineering
Authors
Zuyong He, Xuan Shi, Baozhu Du, Yufeng Qin, Peiqing Cong, Yaosheng Chen,