Abstract
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR-Cas9) system provides a novel genome editing technology that can precisely target a genomic site to disrupt or repair a specific gene. Some CRISPR-Cas9 systems from different bacteria or artificial variants have been discovered or constructed by biologists, and Cas9 nucleases and single guide RNAs (sgRNA) are the major components of the CRISPR-Cas9 system. These Cas9 systems have been extensively applied for identifying therapeutic targets, identifying gene functions, generating animal models, and developing gene therapies. Moreover, CRISPR-Cas9 systems have been used to partially or completely alleviate disease symptoms by mutating or correcting related genes. However, the efficient transfer of CRISPR-Cas9 system into cells and target organs remains a challenge that affects the robust and precise genome editing activity. The current review focuses on delivery systems for Cas9 mRNA, Cas9 protein, or vectors encoding the Cas9 gene and corresponding sgRNA. Non-viral delivery of Cas9 appears to help Cas9 maintain its on-target effect and reduce off-target effects, and viral vectors for sgRNA and donor template can improve the efficacy of genome editing and homology-directed repair. Safe, efficient, and producible delivery systems will promote the application of CRISPR-Cas9 technology in human gene therapy.
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This work was supported by the National Natural and Scientific Foundation of China (81602699 to Zhi-Yao He, 81502677 to Ke Men, 81402302 to Yang Yang), the National High Technology Research and Development Program of China (2015AA020309 to Zhi-Yao He), and the China Postdoctoral Science Foundation Funded Project (2015M570791 to Zhi-Yao He).
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He, ZY., Men, K., Qin, Z. et al. Non-viral and viral delivery systems for CRISPR-Cas9 technology in the biomedical field. Sci. China Life Sci. 60, 458–467 (2017). https://doi.org/10.1007/s11427-017-9033-0
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DOI: https://doi.org/10.1007/s11427-017-9033-0