Recent Advancements in Reducing the Off-Target Effect of CRISPR-Cas9 Genome Editing

Misganaw Asmamaw Mengstie; Muluken Teshome Azezew; Tadesse Asmamaw Dejenie; Assefa Agegnehu Teshome; Fitalew Tadele Admasu; Awgichew Behaile Teklemariam; Anemut Tilahun Mulu; Melaku Mekonnen Agidew; Dagnew Getnet Adugna; Habtamu Geremew; Endeshaw Chekol Abebe

doi:10.2147/BTT.S429411

Recent Advancements in Reducing the Off-Target Effect of CRISPR-Cas9 Genome Editing

Biologics. 2024 Jan 18:18:21-28. doi: 10.2147/BTT.S429411. eCollection 2024.

Affiliations

¹ Department of Biochemistry, College of Medicine and Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia.
² Department of Physiology, College of Medicine and Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia.
³ Department of Medical Biochemistry, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia.
⁴ Department of Anatomy, College of Medicine and Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia.
⁵ Department of Anatomy, School of Medicine, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia.
⁶ College of Health Sciences, Oda Bultum University, Chiro, Ethiopia.

Abstract

The CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)) and the associated protein (Cas9) system, a young but well-studied genome-editing tool, holds plausible solutions to a wide range of genetic disorders. The single-guide RNA (sgRNA) with a 20-base user-defined spacer sequence and the Cas9 endonuclease form the core of the CRISPR-Cas9 system. This sgRNA can direct the Cas9 nuclease to any genomic region that includes a protospacer adjacent motif (PAM) just downstream and matches the spacer sequence. The current challenge in the clinical applications of CRISPR-Cas9 genome-editing technology is the potential off-target effects that can cause DNA cleavage at the incorrect sites. Off-target genome editing confuses and diminishes the therapeutic potential of CRISPR-Cas9 in addition to potentially casting doubt on scientific findings regarding the activities of genes. In this review, we summarize the recent technological advancements in reducing the off-target effect of CRISPR-Cas9 genome editing.

Keywords: CRISPR-Cas9; genome editing; off-target effect; recent advancements; review.

Publication types

Review