Research progress on nucleic acid detection and genome editing of CRISPR/Cas12 system

Abstract

Purpose This work characterizes the applications of CRISPR/Cas12 system, including nucleic acid detection, animal, plant and microbial genome editing. Methods The literature on CRISPR/Cas12 system was collected and reviewed. Results CRISPR/Cas system is an acquired immune system derived from bacteria and archaea, which has become the most popular technology around the world because of its outstanding contribution in genome editing. Type V CRISPR/Cas systems are distinguished by a single RNA-guided RuvC nuclease domain with single effector molecule. Cas12a, the first reported type V CRISPR/Cas system, targets double-stranded DNA (dsDNA) adjacent to PAM sequences and trans-cleaves single-stranded DNA (ssDNA). We present the applications of CRISPR/Cas12 system for nucleic acid detection and genome editing in animals, plants and microorganisms. Furthermore, this review also summarizes the applications of other Cas12 proteins, such as Cas12b, Cas12c, Cas12d, and so on, which further widen the application prospects of CRISPR/Cas12 system. Conclusions Knowledge of the applications of CRISPR/Cas12 system is necessary for improving the understanding of the functional diversity of CRISPR/Cas12 system and also provides significant references for further research and utilization of CRISPR/Cas12 in other new fields. Graphical abstract

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

“…Cas9 and Cas12a, as multi-domain effector proteins with double-leaf structures, offer broad applicability but are reliant on the host cell’s DNA repair mechanism ( Paul and MontoyaCRISPR-Cas12a, 2020 ). In contrast, Cas12b, although smaller in size, demonstrates high specificity, making it invaluable in the genetic engineering ( Yang et al, 2023 ). Cas13 proteins excel in sequence-specific cleavage of single-stranded RNA molecules, enabling precise regulation of RNA products and mitigating unintended pleiotropic effects ( Kavuri et al, 2022 ).…”

Recent advances of biocompatible optical nanobiosensors in liquid biopsy: towards early non-invasive diagnosis

“…2b–g) Revolutionary clustered regularly interspaced short palindromic repeats (CRISPR) technology, specifically CRISPR/Cas12a, has been widely used in gene editing. 31 In addition, circulating tumor (ctDNA), a typical subset of cell-free DNA commonly collected from the blood, is considered an important biomarker for early cancer diagnosis. 32 Huang et al developed a biosensor using nitrophenyl-functionalized black phosphorus nanosheets (NP-BPs) to analyze ctDNA in clinical serum samples within 15 min at an LOD of 50 fM.…”

Point-of-Care Testing for Infectious Diseases Based on Class 2 CRISPR/Cas Technology

“…The CRISPR/Cas12 system has also been applied for gene editing [ 53 , 54 ]. Additionally, it has been successfully used to detect a variety of microorganisms [ 55 , 56 ]. CRISPR/Cas12a-based DETECTR enabled the rapid and specific detection of patient specimens, thereby providing a simple platform for the molecular diagnosis of infectious diseases.…”