Citation: ZHANG Li-Xian, WANG Qi-Fei, CAO Yang, YE Cheng-Yu, CHEN Zhi-Yang, XU Qi-Jie, ZOU Bing-Jie, SONG Qin-Xin, ZHOU Guo-Hua. Enrichment Reporter System of Genome Editing Positive Cells[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(1): 1-12. doi: 10.19756/j.issn.0253-3820.191322 shu

Enrichment Reporter System of Genome Editing Positive Cells

1.
Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing 210009, China;
2.
Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China

Received Date: 6 June 2019
Revised Date: 8 November 2019

Fund Project: This work was supported by the National Natural Science Foundation of China (No.81673390), the Jiangsu Provincial Natural Science Foundation (No. BK20191322), the Jiangsu Provincial Science Fund for Distinguished Young Scholars (No.BK20180005), the Open Project Program of MOE Key Laboratory of Drug Quality Control and Pharmacovigilance (No. DQCP2017MS01), the College Students Innovation Project for the R&D of Novel Drugs (No.J1310032), and the Qing Lan Project of Jiangsu Province.

Genome editing has become a vital tool in medical biology research. The critical mission of facilitating the progress of genome editing is to enrich positive edited cells quickly and effectively. In recent years, researchers have established various reporter systems for selection and enrichment of editing-induced positive cells based on genome repair mechanisms, such as non-homologous end joining, homology directed repair, single strand annealing and inversion, and the principle of the expression of fluorescent protein or resistance tag after genome repair. The T7E1 assay or sequencing method can analyze the mutation of enriched cells with lower background signals and higher mutation ratio. Therefore, these reporter systems can profit the characterization of genome editing effectiveness. Besides, positive cells can be cultured continuously, so this technology possesses a promising prospect of mutated cell line construction and the research of mutated cell functions. This article summarized the design principles and applications of these reporter systems and provided a reference to construct a more perfect evaluating system for genome editing.
- Genome editing,
- Repair mechanism,
- Cell enrichment,
- Reporter system,
- Review
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沈阳化工大学材料科学与工程学院沈阳 110142