AI Article Synopsis
- IGF2 is a crucial growth factor for the development of pigs, and modifying its gene in Lantang pigs via genome editing could enhance lean meat and reduce fat.
- The study compared the effectiveness of two genome editing technologies: CRISPR/Cas9 and zinc finger nucleases (ZFNs), finding that CRISPR/Cas9 was significantly more efficient.
- To boost the percentage of edited cells suitable for further applications, a Single-strand annealing reporter system was employed, which greatly improved the editing efficiency for both CRISPR/Cas9 and ZFNs.
Article Abstract
IGF2 (Insulin-like growth factor 2) is a major growth factor affecting porcine fetal and postnatal development. We propose that the precise modification of IGF2 gene of Chinese indigenous pig breed--Lantang pig by genome editing technology could reduce its backfat thickness, and increase its lean meat content. Here, we tested the genome editing activities of zinc finger nucleases (ZFNs) and CRISPR/Cas9 system on IGF2 gene in the Lantang porcine fetal fibroblasts (PEF). The results indicated that CRISPR/Cas9 presented cutting efficiency up to 9.2%, which was significantly higher than that generated by ZFNs with DNA cutting efficiency lower than 1%. However, even by using CRISPR/Cas9, the relatively lower percentage of genetically modified cells in the transfected population was not satisfied for somatic nuclear transfer (SCNT). Therefore, we used a SSA (Single-strand annealing) reporter system to enrich genetically modified cells induced by ZFN or CRISPR/Cas9. T7 endonuclease I assay revealed that this strategy improved genome editing activity of CRISPR/Cas9 by 5 folds, and was even more effective for improving genome editing efficiency of ZFN.
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| http://dx.doi.org/10.16288/j.yczz.2015.01.008 | DOI Listing |
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