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Current updates on the structural and functional aspects of the CRISPR/Cas13 system for RNA targeting and editing: A
Khaled S Allemailem1, Arshad Husain Rahmani1, Nahlah Makki Almansour2
1Department of Medical Laboratories, College of Applied Medical Sciences, Qassim University, Buraydah 51452, Saudi Arabia.
Abstract:
For centuries, a competitive evolutionary race between prokaryotes and related phages or other mobile genetic elements has led to the diversification of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR‑associated sequence (Cas) genome‑editing systems. Among the different CRISPR/Cas systems, the CRISPR/Cas9 system has been widely studied for its precise DNA manipulation; however, due to certain limitations of direct DNA targeting, off‑target effects and delivery challenges, researchers are looking to perform transient knockdown of gene expression by targeting RNA. In this context, the more recently discovered type VI CRISPR/Cas13 system, a programmable single‑subunit RNA‑guided endonuclease system that has the capacity to target and edit any RNA sequence of interest, has emerged as a powerful platform to modulate gene expression outcomes. All the Cas13 effectors known so far possess two distinct ribonuclease activities. Pre‑CRISPR RNA processing is performed by one RNase activity, whereas the two higher eukaryotes and prokaryotes nucleotide‑binding domains provide the other RNase activity required for target RNA degradation. Recent innovative applications of the type VI CRISPR/Cas13 system in nucleic acid detection, viral interference, transcriptome engineering and RNA imaging hold great promise for disease management. This genome editing system can also be employed by the Specific High Sensitivity Enzymatic Reporter Unlocking platform to identify any tumor DNA. The discovery of this system has added a new dimension to targeting, tracking and editing circulating microRNA/RNA/DNA/cancer proteins for the management of cancer. However, there is still a lack of thorough understanding of the mechanisms underlying some of their functions. The present review summarizes the recent updates on the type VI CRISPR/Cas system in terms of its structural and mechanistic properties and some novel applications of this genome‑editing tool in cancer management. However, some issues, such as collateral degradation of bystander RNA, impose major limitations on its in vivo application. Furthermore, additional challenges and future prospects for this genome editing system are described in the present review.
Insights
The CRISPR/Cas13 system offers precise RNA targeting for gene expression modulation, advancing disease management and cancer research. Further understanding and overcoming limitations like bystander RNA degradation are key for its in vivo applications.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- The evolution of CRISPR-Cas systems, including CRISPR/Cas9, has driven advancements in genome editing.
- Limitations of DNA-targeting CRISPR/Cas9 systems necessitate RNA-targeting alternatives for gene expression modulation.
- The type VI CRISPR/Cas13 system has emerged as a powerful RNA-targeting genome editing tool.
Purpose of the Study:
- To review recent updates on the type VI CRISPR/Cas13 system.
- To summarize its structural and mechanistic properties.
- To highlight novel applications in cancer management and other fields.
Main Methods:
- Review of existing literature on CRISPR/Cas13 systems.
- Analysis of structural and mechanistic properties of Cas13 effectors.
- Compilation of recent applications in nucleic acid detection, viral interference, and cancer management.
Main Results:
- Cas13 systems are programmable RNA-guided endonucleases with distinct ribonuclease activities for RNA processing and degradation.
- Applications include transcriptome engineering, RNA imaging, nucleic acid detection, and viral interference.
- The system shows promise for cancer management through targeting circulating nucleic acids and proteins.
Conclusions:
- The CRISPR/Cas13 system is a versatile platform for RNA manipulation with significant potential in disease management.
- Challenges such as collateral bystander RNA degradation require further investigation for effective in vivo applications.
- Continued research into mechanisms and applications will expand the utility of CRISPR/Cas13 systems.
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