The CRISPR/Cas System: A Customizable Toolbox for Molecular Detection

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated proteins (Cas) are promising molecular diagnostic tools for rapidly and precisely elucidating the structure and function of genomes due to their high specificity, programmability, and multi-system compatibility in nucleic acid recognition. Multiple parameters limit the ability of a CRISPR/Cas system to detect DNA or RNA. Consequently, it must be used in conjunction with other nucleic acid amplification techniques or signal detection techniques, and the reaction components and reaction conditions should be modified and optimized to maximize the detection performance of the CRISPR/Cas system against various targets. As the field continues to develop, CRISPR/Cas systems have the potential to become an ultra-sensitive, convenient, and accurate biosensing platform for the detection of specific target sequences. The design of a molecular detection platform employing the CRISPR/Cas system is asserted on three primary strategies: (1) Performance optimization of the CRISPR/Cas system; (2) enhancement of the detection signal and its interpretation; and (3) compatibility with multiple reaction systems. This article focuses on the molecular characteristics and application value of the CRISPR/Cas system and reviews recent research progress and development direction from the perspectives of principle, performance, and method development challenges to provide a theoretical foundation for the development and application of the CRISPR/CAS system in molecular detection technology.

Adoption of CRISPR-Cas for crop production: present status and future prospects

“…The CRISPR-Cas technology may utilize crops’ natural traits and does not introduce new genes in the event of gene disruption ( Soda, Verma & Giri, 2018 ). Current scientific advancements have shown CRISPR-Cas systems especially type II CRISPR-SpCas9 from Streptococcus pyogenes and type V CRISPR-LbCas12a from Lachnospiraceae bacterium as a versatile technology whose prospects are yet to be thoroughly mined in human biology ( Li et al., 2023a ; Li et al., 2023b ), agriculture, and microbiology ( Brandt & Barrangou, 2019 ; Tomlinson, 2018 ; He et al, 2023 ). With a focus on food security and safety, the effectiveness of CRISPR-Cas systems and other gene editing techniques lies in the ability to keep established croplands productive in the face of a changing climate and conserve the remaining wild areas of the planet.…”

CRISPR-Cas-mediated unfolded protein response control for enhancing plant stress resistance

“…The emergence of CRISPR-Cas technology has revolutionized the study of gene function in plants, as knockout lines generated by CRISPR-Cas tools are more precise and cleaner than traditional methods ( Tien Van et al., 2022 ). The CRISPR-Cas system is highly efficient, customizable, simple, and cost-effective, making it an accessible tool for labs worldwide ( Ahmadzadeh et al., 2019 ; Chen et al., 2019 ; He et al., 2023 ).…”

Advances and Challenges in Cytomegalovirus Detection Methods for Liver Transplant Donors

“…High [87] Abbreviations: NASBA, Nucleic Acid Sequence-Based Amplification; LAMP, Loop-Mediated Isothermal Amplification; DNA-RNA, Deoxyribonucleic Acid-Ribonucleic Acid. CMV, Cytomegalovirus; CRISPR-Cas, Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-Associated.…”