CRISPR/Cas technology as a promising weapon to combat viral infections

Abstract

The versatile clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system has emerged as a promising technology for therapy and molecular diagnosis. It is especially suited for overcoming viral infections outbreaks, since their effective control relies on an efficient treatment, but also on a fast diagnosis to prevent disease dissemination. The CRISPR toolbox offers DNA‐ and RNA‐targeting nucleases that constitute dual weapons against viruses. They allow both the manipulation of viral and host genomes for therapeutic purposes and the detection of viral nucleic acids in “Point of Care” sensor devices. Here, we thoroughly review recent advances in the use of the CRISPR/Cas system for the treatment and diagnosis of viral deleterious infections such as HIV or SARS‐CoV‐2, examining their strengths and limitations. We describe the main points to consider when designing CRISPR antiviral strategies and the scientific efforts to develop more sensitive CRISPR‐based viral detectors. Finally, we discuss future prospects to improve both applications. Also see the video abstract here: https://www.youtube.com/watch?v=C0z1dLpJWl4

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

“…The main focus of CRISPR-based biosensing techniques has been the diagnostics of pathogen infection benefits from their DNA- and RNA-targeting nucleases ( Escalona-Noguero et al, 2021 ). Amid the ongoing pandemic of COVID-19, enormous efforts have been paid for the detection of SARS-CoV-2 ( Nouri et al, 2021 ).…”

“…Currently, clustered regularly interspaced short palindromic repeats (CRISPR) based biosensors have been developed as powerful tools for nucleic acid sensing and widely applied to the rapid diagnosis of infectious pathogens ( Kellner et al, 2019 ; Qin et al, 2019 ; Broughton et al, 2020 ; Ding R. et al, 2021 ; Song F. et al, 2021 ; Chen et al, 2021c ; Escalona-Noguero et al, 2021 ; Ge et al, 2021 ) and the detection of DNA or miRNAs associated with diseases such as cancer ( Li B. et al, 2021 ; Wang et al, 2021c ; Chen Y. et al, 2021 ). These biosensors rely on the CRISPR systems containing CRISPR-associated proteins (Cas) with nonspecific endonuclease activity to efficiently cleave specific targets via guide RNAs (gRNAs) ( Cong et al, 2013 ; Mali et al, 2013 ; van der Oost, 2013 ).…”

The Challenge of CRISPR-Cas Toward Bioethics

“…In addition to the above, there is the possibility of interrupting the advance of bacterial resistance to antibiotics, in addition to decreasing the virulence of bacterial isolates that cause infections. The purpose is to always try to help human health, but care must be taken not to cause unwanted alterations in the patient (Escalona-Noguero et al, 2021;Tsou et al, 2021;Yadav et al, 2021).…”

CASCADE: Naked eye-detection of SARS-CoV-2 using Cas13a and gold nanoparticles

“…Following this step, amplification products must be specifically detected. Some of the most successful detection methods are based on the CRISPR/Cas system, which can be easily coupled to isothermal amplification strategies [ 5 ]. The Class 2 CRISPR/Cas system consists of a Cas nuclease forming a ribonucleoprotein (RNP) complex with a guide RNA (crRNA) which is complementary to a target nucleic acid (DNA/RNA).…”

“…So far, the combination of Cas13 and AuNPs for the naked-eye detection of SARS-CoV-2 RNA remains an unexplored field. Cas13a has been successfully used to detect viral nucleic acids using fluorimetric and lateral flow read-outs [ 5 ] and it is a good candidate for naked-eye sensing. While Cas12a target recognition relies on the presence of a protospacer adjacent motif (PAM), Cas13 lacks this kind of sequence requirements, which makes it more versatile.…”