Page 90 - ebook

[A. Biochemistry/Molecular Biology] A-45

Transcriptome-wide off-target analysis of RNA-targeting

type-VI CRISPR system

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Seong-Ho Park¹ , Seokju Park⁴ , Jin Wu Nam⁴,⁵,⁶*, Junho K Hur¹,³*
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¹Department of Medicine, Major in Medical Genetics, Graduate School, Hanyang University, Seoul 04736, Korea,

²Graduate School of Biomedical Science & Engineering, Hanyang University, Seoul 04736, Korea, ³Department of

Genetics, College of Medicine, Hanyang University, Seoul 04736, Korea, ⁴Department of Life Sciences, College of
Natural Science, Hanyang University, Seoul 04736, Korea, ⁵Research Institute for Natural Sciences, Hanyang
University, Seoul 04736, Korea

The CRISPR/Cas systems are known as the 3rd generation gene-editing tool, and have been mostly applied for
DNA editing. CRISPR/Cas systems are widely studied in biological and medical research fields as the CRISPR systems

can conduct direct editing of DNA to permanently change the genome. However, one o the safety concerns of the
CISPR/Cas systems have been off-target effects, that induces unanticipated DNA sequence changes in the genome.

Many studies have been trying to overcome the off-target issue by developing highly precise CRISPR/Cas systems
for gene therapy. To this end, CRISPR-Cas13, an RNA editing CRISPR system, had been studied as a tool for targeted

gene regulation without DNA changes. However, the Cas13 had not widely used research because of low RNA
editing efficiency. According to a recent study, the CasRX, a type of Cas13d, was high efficiency and small size, it

was an advantage in virus loading. In this study, we performed that degradome-seq for detected of the cleavage
site in RNA for observed the off-target effect, the disadvantage of the CRISPR/Cas system, in CasRX. And we
researched guide RNA screening for optimized gRNA design.