Zinc Finger Proteins

Jiani Xing, Cunfang Zhang, Kun Xu, Linyong Hu, Ling Wang, Tingting Zhang, Chonghua Ren, Zhiying Zhang, Xing, Jiani, Zhang, Cunfang, Xu, Kun, Hu, Linyong, Wang, Ling, Zhang, Tingting, Ren, Chonghua, Zhang, Zhiying

Using engineered nucleases such as zinc-finger nucleases (ZFNs) and TALE nuclease (TALEN) to accomplish genome editing often causes high cellular toxicity because of the consistent expression of artificial nucleases and off-targeting effect. And lacking selection marker in modified cells makes it hard to enrich these positive cells. Here we introduce a method by incorporating a surrogate reporter enrichment into a suicidal ZFN system, which is designed by a pair of ZFN expression cassettes flanked with its target sites. Our data demonstrated that this modified system achieved almost the same ZFN activity as the original method but reduced ~40% toxicity. This new suicidal ZFN expression system coupled with a surrogate reporter not only enables decreased cellular toxicity but also makes the genetic modified cells to be enriched by EGFP analysis.