Principles and Applications of CRISPR Toolkit in Virus Manipulation, Diagnosis, and Virus-Host Interactions

Abstract

Viruses are one of the most important concerns for human health, and overcoming viral infections is a worldwide challenge. However, researchers have been trying to manipulate viral genomes to overcome various disorders, including cancer, for vaccine development purposes. CRISPR (clustered regularly interspaced short palindromic repeats) is becoming one of the most functional and widely used tools for RNA and DNA manipulation in multiple organisms. This approach has provided an unprecedented opportunity for creating simple, inexpensive, specific, targeted, accurate, and practical manipulations of viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), human immunodeficiency virus-1 (HIV-1), and vaccinia virus. Furthermore, this method can be used to make an effective and precise diagnosis of viral infections. Nevertheless, a valid and scientifically designed CRISPR system is critical to make more effective and accurate changes in viruses. In this review, we have focused on the best and the most effective ways to design sgRNA, gene knock-in(s), and gene knock-out(s) for virus-targeted manipulation. Furthermore, we have emphasized the application of CRISPR technology in virus diagnosis and in finding significant genes involved in virus-host interactions.

Insertion-and-Deletion Mutations between the Genomes of SARS-CoV, SARS-CoV-2, and Bat Coronavirus RaTG13

“…The CRISPR-Cas system is useful in RNA and DNA editing that realized relatively precise genome modifications, including an induction of indel to the DNA sequences ( 44 , 45 ). The CRISPR toolkit is further expected to shorten the time for developing an effective live attenuated vaccine for some viruses or realizing viral load reduction after viral infections ( 45 , 46 ). However, whether the CRISPR-Cas13 system (which targets RNA and not DNA) can realize the long insertion-and-deletion mutations in RNA molecules that were observed in the present study remains unknown.…”

Research and Therapeutic Approaches in Stem Cell Genome Editing by CRISPR Toolkit

“…Some different stem cell types have so far been successfully used in clinical studies and have received scientific approval. Besides viral genome modification, more and more publications are confirming that CRISPR/Cas9 genome editing is a potent technique that can greatly advance biomedicine, such as in virus genome editing and in stem cell research [ 11 , 17 , 18 , 19 ]. Organoids are interesting new systems that help us improve our knowledge of diseases’ mechanisms, development, evolution, homeostasis, and therapy.…”