CRISPR-Cas13 and Its Impact on Biology and Biomedicine
The ground-breaking CRISPR-Cas13 system has become a potent tool for RNA modification in the rapidly changing field of genetic engineering, leading to new possibilities in biological and biomedical applications. This programmable system offers a flexible platform for applications like RNA editing, disease treatment, and interference with viral infections, in addition to enabling targeted RNA cleavage.
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Knowing the CRISPR-Cas13 System:
The CRISPR-Cas13 system, known for its ability to cleave both target and non-target RNA simultaneously, has found widespread applications in various fields. This dual cleavage capability has been harnessed for RNA knockout strategies, disease treatment, interference with viral infections, screening of loss-of-function mutants, and molecular detection of biological agents.
RNA Editing with CRISPR-Cas13:
One of the key advancements facilitated by CRISPR-Cas13 is RNA editing. By engineering RNA-targeting Cas effectors to retain their binding ability to target RNA in a controlled manner, researchers can now remove nuclease activity and fuse Cas13 with ADAR (adenosine deaminase acting on RNA). This programmable RNA editing approach enables the rewriting of gene expression without altering genome sequences.
Challenges in RNA Editing:
While the potential of RNA editing is immense, current tools for specific RNA editing face challenges, such as the assembly of guide RNA into an RNA/protein complex, causing delivery barriers and low editing efficiency. The CRISPR/Cas13 system addresses these challenges, offering a more efficient and precise solution for RNA editing.
The Adaptive Immune System of CRISPR/Cas13:
Originally functioning as an ‘adaptive’ immune system in bacteria and archaea, the CRISPR/Cas13 system acts as a defense mechanism against invading RNA elements, including RNA viruses. By recognizing and degrading invading RNAs, this system has proven instrumental in molecular defense strategies.
Historical Context: RNA Editing Discovery:
In 1986, Benne published groundbreaking work on RNA editing, introducing a process that precisely inserts four uridines at a defined position in the Cox2 transcript. This discovery paved the way for understanding RNA editing and its implications in gene expression, providing a foundation for future research.
CRISPR/Cas13 in Plant Transcriptomics:
The application of CRISPR/Cas13 in plants has offered valuable insights into plant transcriptomic studies. Beyond imparting tolerance against plant viruses, CRISPR/Cas13 is emerging as a powerful tool for targeting mRNA, circRNA, and other non-coding RNAs, expanding its potential impact on plant genetics.
Cas13 vs. Cas9 and Cas12a:
While Cas9 and Cas12a are renowned for DNA manipulation, the Cas13 family stands out for its ability to target RNA sequences. This becomes particularly significant in antiviral therapy, where viruses may possess RNA or DNA as genetic material. Cas12a and Cas9 remain effective against DNA viruses and RNA viruses with dsDNA intermediaries.
RNA Editing and Disease Treatment:
The ability to correct disease-causing mutations through efficient and precise recoding represents a primary goal of RNA editing. Recent studies have linked RNA editing to cancer development, offering potential avenues for cancer diagnosis and treatment. By understanding and manipulating RNA sequences, researchers are uncovering new possibilities for combating diseases at the genetic level.
Types of RNA Editing:
RNA editing involves the post-transcriptional insertion, deletion, or modification of nucleotides. Notably, it excludes modifications made to primary transcripts in RNA processing. In higher eukaryotes, A-to-I base modifications are the most common form of RNA editing, occurring in mRNAs, tRNAs, and rRNAs across various cellular compartments.
The integration of CRISPR-Cas13 into the realm of RNA modification represents a groundbreaking achievement in genetic engineering. From its origins as a bacterial defense mechanism to its applications in RNA editing and disease treatment, CRISPR-Cas13 has reshaped our approach to manipulating genetic information. As research progresses, the potential for this programmable system to revolutionize biological and biomedical applications continues to expand, offering hope for innovative solutions in the ongoing quest to understand and harness the power of genetic code.
References:
Abbott T. R., Dhamdhere G., Liu Y., Lin X., Goudy L., Zeng L., et al. (2020). Development of CRISPR as an antiviral strategy to combat SARS-CoV-2 and influenza. Cell 181 865–876.e12. – PMC – PubMed
Abudayyeh O. O., Gootenberg J. S., Essletzbichler P., Han S., Joung J., Belanto J. J., et al. (2017). RNA targeting with CRISPR-Cas13. Nature 550 280–284. – PMC – PubMed
Abudayyeh O. O., Gootenberg J. S., Franklin B., Koob J., Kellner M. J., Ladha A., et al. (2019). A cytosine deaminase for programmable single-base RNA editing. Science 365 382–386. 10.1126/science.aax7063 – DOI – PMC – PubMed
Abudayyeh O. O., Gootenberg J. S., Konermann S., Joung J., Slaymaker I. M., Cox D. B., et al. (2016). C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science 353:aaf5573. 10.1126/science.aaf5573 – DOI – PMC – PubMed
Ackerman C. M., Myhrvold C., Thakku S. G., Freije C. A., Metsky H. C., Yang D. K., et al. (2020). Massively multiplexed nucleic acid detection with Cas13. Nature 582 277–282. 10.1038/s41586-020-2279-8 – DOI – PMC – PubMed
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