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Cited by 430CRISPR and Genetic EngineeringRNA Interference and Gene DeliveryNanoparticle-Based Drug Delivery

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

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Zhang, Song, Shen, Jiangtao, Li, Dali, Cheng, Yiyun·2021·Theranostics
DOI 10.7150/thno.47007PMC PMC7738854
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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.

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  • RNP methods show a 2.5- to 28-fold reduction in off-target effects compared to plasmid-based methods.

    “More specifically, a 2.5- to 28-fold reduction in off-target effects has been reported with an RNP method compared to plasmid-based methods (Liang et al., 2015; Zhang et al., 2021).”

    From Efficient genetic editing of human intestinal organoids using ribonucleoprotein-based CRISPR (2023) · cites this paper as Zhang et al., 2021

    The Conclusions and perspectives section says direct RNP delivery has lower off-target effects than Cas9 plasmid delivery. It does not report the claimed 2.5- to 28-fold reduction, so the quantitative part is unsupported by this source. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

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200 of 254 shown

Cited by

Sorted by citations

Authors
Zhang, S., Shen, J., Li, D., & Cheng, Y.
Affiliations
  • South China University of Technology (CN)
  • Taizhou Second People's Hospital (CN)
  • East China Normal University (CN)
Corresponding author
  • Li, Dali
  • Cheng, Yiyun
ORCID
Venue
Theranostics
Publisher
Ivyspring International Publisher
Published
2021
Volume
11
Issue
2
Pages
614-648
ISSN
1838-7640 (ISSN-L)
DOI
10.7150/thno.47007
PMC
PMC7738854
OpenAlex
W3097071384
Cited by
430
References
19
Citations by year
2026: 60 · 2025: 90 · 2024: 89 · 2023: 71 · 2022: 81 · 2021: 35 · 2020: 3
FWCI
15.41
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Theranostics (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
CRISPR/Cas9 genome editing, single guide RNA, stimuli-responsive carriers, targeted delivery, intracellular delivery, off-target effects, gene therapy
MeSH terms
Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Animals, Humans, RNA, Guide, Kinetoplastida, Gene Transfer Techniques, CRISPR-Cas Systems

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