Molecular Switch Engineering for Precise Genome Editing

Abstract

Genome editing technology commenced in 1996 with the discovery of the first zinc-finger nuclease. Application of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) associated protein 9 (Cas9) technology to genome editing of mammalian cells allowed researchers to use genome editing more easily and cost-effectively. However, one of the technological problems that remains to be solved is “off-target effects”, which are unexpected mutations in nontarget DNA. One significant improvement in genome editing technology has been achieved with molecular/protein engineering. The key to this engineering is a “switch” to control function. In this review, we discuss recent efforts to design novel “switching” systems for precise editing using genome editing tools.

Molecular evolution and functional modification of plant miRNAs with CRISPR

“…A recently developed synthetic approach of crRNA/tracrRNA duplex (gRNA) with photocaging groups can avoid and separate the gRNA from recognizing and combining with the targeted sequence [131]. This approach relies on a solid-phase synthesis of the photocaged gRNAs, with simpler purification and characterization processes in comparison with engineering a light-responsive protein [132,133]. A good example was reported in zebrafish (Danio rerio), where this approach was applied to spatiotemporally control gene editing, enabling the use of this strategy for developmental biology and tissue engineering applications [131].…”