Engineering the Delivery System for CRISPR-Based Genome Editing

Overview

Journal Trends Biotechnol

Specialty Biochemistry

Date 2018 Jan 7

PMID 29305085

Citations 142

Authors

Zachary Glass

Matthew Lee

Yamin Li

Qiaobing Xu

Affiliations

Soon will be listed here.

Abstract

Clustered regularly interspaced short palindromic repeat-CRISPR-associated protein (CRISPR-Cas) systems, found in nature as microbial adaptive immune systems, have been repurposed into an important tool in biological engineering and genome editing, providing a programmable platform for precision gene targeting. These tools have immense promise as therapeutics that could potentially correct disease-causing mutations. However, CRISPR-Cas gene editing components must be transported directly to the nucleus of targeted cells to exert a therapeutic effect. Thus, efficient methods of delivery will be critical to the success of therapeutic genome editing applications. Here, we review current strategies available for in vivo delivery of CRISPR-Cas gene editing components and outline challenges that need to be addressed before this powerful tool can be deployed in the clinic.

Citing Articles

From Origin to the Present: Establishment, Mechanism, Evolutions and Biomedical Applications of the CRISPR/Cas-Based Macromolecular System in Brief.

Yuan Z Molecules. 2025; 30(4).

PMID: 40005257 PMC: 11858448. DOI: 10.3390/molecules30040947.

Harnessing nanotechnology for cancer treatment.

Zhu J, Lee H, Huang R, Zhou J, Zhang J, Yang X Front Bioeng Biotechnol. 2025; 12:1514890.

PMID: 39902172 PMC: 11788409. DOI: 10.3389/fbioe.2024.1514890.

Genetic enhancement: an avenue to combat aging-related diseases.

Cai Y, Ji Z, Wang S, Zhang W, Qu J, Belmonte J Life Med. 2025; 1(3):307-318.

PMID: 39872744 PMC: 11749557. DOI: 10.1093/lifemedi/lnac054.

The role of nanoparticles in transforming plant genetic engineering: advancements, challenges and future prospects.

Rani N, Kumari K, Hooda V Funct Integr Genomics. 2025; 25(1):23.

PMID: 39841261 DOI: 10.1007/s10142-025-01528-x.

Improving the use of CRISPR/Cas9 gene editing machinery as a cancer therapeutic tool with the help of nanomedicine.

Fatima H, Singh D, Muhammad H, Acharya S, Aziz M 3 Biotech. 2024; 15(1):17.

PMID: 39711922 PMC: 11656010. DOI: 10.1007/s13205-024-04186-1.

References

Hashimoto M, Takemoto T . Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing. Sci Rep. 2015; 5:11315. PMC: 4463957. DOI: 10.1038/srep11315. View

Zetsche B, Gootenberg J, Abudayyeh O, Slaymaker I, Makarova K, Essletzbichler P . Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell. 2015; 163(3):759-71. PMC: 4638220. DOI: 10.1016/j.cell.2015.09.038. View

Sternberg S, Doudna J . Expanding the Biologist's Toolkit with CRISPR-Cas9. Mol Cell. 2015; 58(4):568-74. DOI: 10.1016/j.molcel.2015.02.032. View

Kim S, Kim D, Cho S, Kim J, Kim J . Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 2014; 24(6):1012-9. PMC: 4032847. DOI: 10.1101/gr.171322.113. View

Horvath P, Barrangou R . CRISPR/Cas, the immune system of bacteria and archaea. Science. 2010; 327(5962):167-70. DOI: 10.1126/science.1179555. View

Bazak R, Houri M, El Achy S, Kamel S, Refaat T . Cancer active targeting by nanoparticles: a comprehensive review of literature. J Cancer Res Clin Oncol. 2014; 141(5):769-84. PMC: 4710367. DOI: 10.1007/s00432-014-1767-3. View

Lee K, Conboy M, Park H, Jiang F, Kim H, DeWitt M . Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA induces homology-directed DNA repair. Nat Biomed Eng. 2018; 1:889-901. PMC: 5968829. DOI: 10.1038/s41551-017-0137-2. View

Liang X, Potter J, Kumar S, Zou Y, Quintanilla R, Sridharan M . Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J Biotechnol. 2015; 208:44-53. DOI: 10.1016/j.jbiotec.2015.04.024. View

Hattori Y, Suzuki S, Kawakami S, Yamashita F, Hashida M . The role of dioleoylphosphatidylethanolamine (DOPE) in targeted gene delivery with mannosylated cationic liposomes via intravenous route. J Control Release. 2005; 108(2-3):484-95. DOI: 10.1016/j.jconrel.2005.08.012. View

10.

Wang M, Glass Z, Xu Q . Non-viral delivery of genome-editing nucleases for gene therapy. Gene Ther. 2016; 24(3):144-150. DOI: 10.1038/gt.2016.72. View

11.

Urnov F, Miller J, Lee Y, Beausejour C, Rock J, Augustus S . Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature. 2005; 435(7042):646-51. DOI: 10.1038/nature03556. View

12.

Kim D, Kim J, Hur J, Been K, Yoon S, Kim J . Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016; 34(8):863-8. DOI: 10.1038/nbt.3609. View

13.

Onuki Y, Obata Y, Kawano K, Sano H, Matsumoto R, Hayashi Y . Membrane Microdomain Structures of Liposomes and Their Contribution to the Cellular Uptake Efficiency into HeLa Cells. Mol Pharm. 2015; 13(2):369-78. DOI: 10.1021/acs.molpharmaceut.5b00601. View

14.

Bonamassa B, Hai L, Liu D . Hydrodynamic gene delivery and its applications in pharmaceutical research. Pharm Res. 2010; 28(4):694-701. PMC: 3064722. DOI: 10.1007/s11095-010-0338-9. View

15.

Hughes T, Langer S, Virtanen S, Chavez R, Watkins L, Milligan E . Immunogenicity of intrathecal plasmid gene delivery: cytokine release and effects on transgene expression. J Gene Med. 2009; 11(9):782-90. PMC: 3649870. DOI: 10.1002/jgm.1364. View

16.

Kamimura K, Yokoo T, Abe H, Kobayashi Y, Ogawa K, Shinagawa Y . Image-Guided Hydrodynamic Gene Delivery: Current Status and Future Directions. Pharmaceutics. 2015; 7(3):213-23. PMC: 4588196. DOI: 10.3390/pharmaceutics7030213. View

17.

Ran F, Cong L, Yan W, Scott D, Gootenberg J, Kriz A . In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015; 520(7546):186-91. PMC: 4393360. DOI: 10.1038/nature14299. View

18.

Komor A, Kim Y, Packer M, Zuris J, Liu D . Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature. 2016; 533(7603):420-4. PMC: 4873371. DOI: 10.1038/nature17946. View

19.

Prabhakar U, Maeda H, Jain R, Sevick-Muraca E, Zamboni W, Farokhzad O . Challenges and key considerations of the enhanced permeability and retention effect for nanomedicine drug delivery in oncology. Cancer Res. 2013; 73(8):2412-7. PMC: 3916009. DOI: 10.1158/0008-5472.CAN-12-4561. View

20.

Tang L, Zeng Y, Du H, Gong M, Peng J, Zhang B . CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein. Mol Genet Genomics. 2017; 292(3):525-533. DOI: 10.1007/s00438-017-1299-z. View