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Theranostics 2021; 11(2):614-648. doi:10.7150/thno.47007 This issue Cite

Review

Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing

Song Zhang¹, Jiangtao Shen², Dali Li^3✉, Yiyun Cheng^1,3✉

1. South China Advanced Institute for Soft Matter Science and Technology, School of Molecular Science and Engineering, South China University of Technology, Guangzhou 510640, China.
2. The Second People's Hospital of Taizhou affiliated to Yangzhou University, Taizhou, 225500, China.
3. Shanghai Key Laboratory of Regulatory Biology, East China Normal University, Shanghai 200241, China.

✉ Corresponding authors: yychengustc.edu.cn; dlliecnu.edu.cn

Abstract

CRISPR/Cas9 genome editing has gained rapidly increasing attentions in recent years, however, the translation of this biotechnology into therapy has been hindered by efficient delivery of CRISPR/Cas9 materials into target cells. Direct delivery of CRISPR/Cas9 system as a ribonucleoprotein (RNP) complex consisting of Cas9 protein and single guide RNA (sgRNA) has emerged as a powerful and widespread method for genome editing due to its advantages of transient genome editing and reduced off-target effects. In this review, we summarized the current Cas9 RNP delivery systems including physical approaches and synthetic carriers. The mechanisms and beneficial roles of these strategies in intracellular Cas9 RNP delivery were reviewed. Examples in the development of stimuli-responsive and targeted carriers for RNP delivery are highlighted. Finally, the challenges of current Cas9 RNP delivery systems and perspectives in rational design of next generation materials for this promising field will be discussed.

Keywords: genome editing, CRISPR, RNP, polymers, nanoparticles

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