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Engineering the Delivery System for CRISPR-Based Genome Editing.
Literature Information
| DOI | 10.1016/j.tibtech.2017.11.006 |
|---|---|
| PMID | 29305085 |
| Journal | Trends in biotechnology |
| Impact Factor | 14.9 |
| JCR Quartile | Q1 |
| Publication Year | 2018 |
| Times Cited | 155 |
| Keywords | CRISPR, Cas9, clinical, delivery, gene editing |
| Literature Type | Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S., Review |
| ISSN | 0167-7799 |
| Pages | 173-185 |
| Issue | 36(2) |
| Authors | Zachary Glass, Matthew Lee, Yamin Li, Qiaobing Xu |
TL;DR
This paper reviews the current strategies for the in vivo delivery of CRISPR-Cas gene editing components, emphasizing the importance of efficient delivery methods for the therapeutic potential of these tools in correcting disease-causing mutations. The discussion highlights existing challenges that must be overcome to enable the clinical application of CRISPR-Cas technologies in genome editing.
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CRISPR · Cas9 · clinical · delivery · gene editing
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.
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Primary Questions Addressed
- What are the most promising delivery methods currently being researched for CRISPR-Cas components in vivo?
- How do different delivery systems impact the efficiency and specificity of CRISPR gene editing in various cell types?
- What are the key challenges faced in the development of safe and effective delivery systems for CRISPR therapeutics?
- How can nanotechnology be integrated into CRISPR delivery systems to enhance targeting and reduce off-target effects?
- What regulatory considerations must be addressed when developing delivery systems for CRISPR-based therapies in clinical settings?
Key Findings
Research Background and Purpose
CRISPR-Cas systems, originally part of microbial adaptive immune systems, have been transformed into a powerful tool for genome editing. The ability to program these systems for precise gene targeting holds great promise for therapeutic applications, particularly in correcting disease-causing mutations. However, effective delivery of CRISPR components to the target cells' nuclei remains a significant challenge that must be addressed for successful clinical applications.
Main Methods/Materials/Experimental Design
The review discusses various strategies for in vivo delivery of CRISPR-Cas gene editing components, including:
- Viral Delivery: Utilizing engineered viruses like adeno-associated virus (AAV) to deliver Cas9 and guide RNA (gRNA).
- Nonviral Delivery: Methods such as electroporation, hydrodynamic injection, and microinjection.
- Chemical Delivery: Using lipid nanoparticles, gold nanoparticles, and cell-penetrating peptides (CPPs) to enhance delivery efficiency.
Key Results and Findings
- Delivery Formats: Cas9 can be delivered as DNA, mRNA, or protein, each with unique advantages and challenges:
- DNA Delivery: Offers stable expression but slower onset of gene editing.
- mRNA Delivery: Provides quicker onset but is transient and susceptible to degradation.
- Protein Delivery: Enables immediate therapeutic action but faces significant delivery hurdles.
- Delivery Vehicles: A variety of vehicles, both viral and nonviral, have been explored for their efficacy in delivering CRISPR components, each with distinct pros and cons related to specificity, efficiency, and safety.
Main Conclusions/Significance/Innovativeness
The review highlights the critical importance of developing effective delivery systems for CRISPR technology to maximize its therapeutic potential. It underscores the need for tailored delivery strategies to enhance specificity and minimize off-target effects, which are major concerns in clinical applications. Advances in delivery vehicles and methods will be crucial for the future of CRISPR-based therapies, particularly as new applications like base editing and homology-directed repair emerge.
Research Limitations and Future Directions
- Limitations: The review acknowledges that many delivery methods are still in experimental stages and may not yet be suitable for human application. Issues such as immunogenicity, off-target effects, and the complexity of delivery to specific tissues need further investigation.
- Future Directions: The field is rapidly evolving, with ongoing research into novel delivery systems and CRISPR applications. Future studies should focus on enhancing the specificity and efficiency of delivery methods while exploring new therapeutic uses of CRISPR technology, including combating viral infections and personalized medicine approaches.
Summary Table of Delivery Methods
| Delivery Method | Advantages | Disadvantages |
|---|---|---|
| Viral (AAV) | High efficiency, targeted delivery | Risk of insertional mutagenesis, size limitations |
| Electroporation | Effective for large molecules | Not suitable for therapeutic use in humans |
| Hydrodynamic Injection | Good tissue distribution | Limited to small animal models |
| Microinjection | High efficiency, precise dosage | Impractical for high-throughput applications |
| Lipid Nanoparticles | FDA-approved, versatile | Potential toxicity, immunogenic response |
| Gold Nanoparticles | Stable, promising for in vivo delivery | Limited research in clinical settings |
| CPPs | Enhances membrane penetration | Lack of specificity, no degradation protection |
This structured summary encapsulates the critical aspects of the review, highlighting the advancements and ongoing challenges in CRISPR delivery systems.
References
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- The role of dioleoylphosphatidylethanolamine (DOPE) in targeted gene delivery with mannosylated cationic liposomes via intravenous route. - Yoshiyuki Hattori;Sachiko Suzuki;Shigeru Kawakami;Fumiyoshi Yamashita;Mitsuru Hashida - Journal of controlled release : official journal of the Controlled Release Society (2005)
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Literatures Citing This Work
- Fshb Knockout Mouse Model, Two Decades Later and Into the Future. - T Rajendra Kumar - Endocrinology (2018)
- Role of Gene Therapy in Pancreatic Cancer-A Review. - Mizuho Sato-Dahlman;Keith Wirth;Masato Yamamoto - Cancers (2018)
- Non-viral delivery systems for CRISPR/Cas9-based genome editing: Challenges and opportunities. - Ling Li;Shuo Hu;Xiaoyuan Chen - Biomaterials (2018)
- Gene editing in the context of an increasingly complex genome. - K Blighe;L DeDionisio;K A Christie;B Chawes;S Shareef;T Kakouli-Duarte;C Chao-Shern;V Harding;R S Kelly;L Castellano;J Stebbing;J A Lasky-Su;M A Nesbit;C B T Moore - BMC genomics (2018)
- CRISPR-Cas guides the future of genetic engineering. - Gavin J Knott;Jennifer A Doudna - Science (New York, N.Y.) (2018)
- CRISPR-delivery particles targeting nuclear receptor-interacting protein 1 (Nrip1) in adipose cells to enhance energy expenditure. - Yuefei Shen;Jessica L Cohen;Sarah M Nicoloro;Mark Kelly;Batuhan Yenilmez;Felipe Henriques;Emmanouela Tsagkaraki;Yvonne J K Edwards;Xiaodi Hu;Randall H Friedline;Jason K Kim;Michael P Czech - The Journal of biological chemistry (2018)
- DNA, RNA, and Protein Tools for Editing the Genetic Information in Human Cells. - Xiaoyu Chen;Manuel A F V Gonçalves - iScience (2018)
- Transfection by cationic gemini lipids and surfactants. - M Damen;A J J Groenen;S F M van Dongen;R J M Nolte;B J Scholte;M C Feiters - MedChemComm (2018)
- Nanoparticle-Based Delivery of CRISPR/Cas9 Genome-Editing Therapeutics. - Brittany E Givens;Youssef W Naguib;Sean M Geary;Eric J Devor;Aliasger K Salem - The AAPS journal (2018)
- A peptide delivery system sneaks CRISPR into cells. - Xingang Guan;Zhimin Luo;Wujin Sun - The Journal of biological chemistry (2018)
... (145 more literatures)
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