CRISPR/Cas13 — A New Way to Study RNA    

The CRISPR/Cas technology "innovated" our ability to accurately edit genes and target gene expression, which has been rapidly utilized to perform a variety of genome engineering tasks. Recently, a novel RNA-targeting CRISPR effector protein called Cas13 has been demonstrated which is similar to the CRISPR/Cas9 system. However, unlike Cas9, which targets DNA, Cas13 targets and cleaves single-stranded RNA. This represents a potentially safe alternative to DNase Cas9 because it induces a loss of function phenotype without the genomic change.

Cas13 was first discovered in L. shahii, a species of ciliate, while researchers are looking for previously unknown CRISPR systems. To date, four Cas13 protein families have been identified: Cas13a, originally known as Class 2 candidate protein 2 (C2c2), Cas13b, Cas13c and Cas13d. Cas13 contains two higher eukaryotic and prokaryotic nucleotide binding (HEPN) domains that together form a ribonuclease active site that enables it to target CRISPR effectors as RNA-directed RNA.

The known Cas13a, Cas13d effector are capable of efficiently cleaving complementary target single-stranded RNA. By improving the Cas13 effector function, a system called REPAIR is developed to edit full-length transcripts containing pathogen mutations, providing a broad opportunity for accurate base editing. In addition, members of Cas13, Cas13a and Cas13b are being developed for the correction of therapeutic genes at the RNA level and for detection of viral pathogens. All of these features give us an insight into the potential of the CRISPR toolkit for precision transcriptome engineering and may lead to the expansion of CRISPR technology in cancer treatment and diagnosis. Although the potential clinical application is enormous, overcoming transmission barriers remains critical to the clinical success of the CRISPR/Cas13 system or any other nucleic acid-based therapeutic strategy.

Figure 1: CRISPR/Cas13, as a versatile toolkit for transcriptome engineering (Javier T, 2018)

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