CRISPR/Cas9 therapeutics: progress and prospects

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene-editing technology is the ideal tool of the future for treating diseases by permanently correcting deleterious base mutations or disrupting disease-causing genes with great precision and efficiency. A variety of efficient Cas9 variants and derivatives have been developed to cope with the complex genomic changes that occur during diseases. However, strategies to effectively deliver the CRISPR system to diseased cells in vivo are currently lacking, and nonviral vectors with target recognition functions may be the focus of future research. Pathological and physiological changes resulting from disease onset are expected to serve as identifying factors for targeted delivery or targets for gene editing. Diseases are both varied and complex, and the choice of appropriate gene-editing methods and delivery vectors for different diseases is important. Meanwhile, there are still many potential challenges identified when targeting delivery of CRISPR/Cas9 technology for disease treatment. This paper reviews the current developments in three aspects, namely, gene-editing type, delivery vector, and disease characteristics. Additionally, this paper summarizes successful examples of clinical trials and finally describes possible problems associated with current CRISPR applications.

Gene therapy for pediatric genetic kidney diseases

“…CRISPR/Cas9 is an RNA-guided DNA nuclease system that has evolved as an adaptive immune mechanism in prokaryotes. 9 Typically, the CRISPR/Cas9 system is composed of a single-stranded RNA (sgRNA) and Cas9 protein. The sgRNA directs the nuclease to a specific DNA site, making the system modular and easy to engineer.…”

“…CRISPR/Cas9 is an RNA‐guided DNA nuclease system that has evolved as an adaptive immune mechanism in prokaryotes 9 . Typically, the CRISPR/Cas9 system is composed of a single‐stranded RNA (sgRNA) and Cas9 protein.…”

“…Gene editing is a molecular technology that involves deleting, inserting, or replacing a specific segment or specific nucleotides of the genome to induce specific changes in the genome. Over the last 2 decades, the field of gene editing has witnessed remarkable advancements with the emergence of various technologies, such as meganucleases, zinc finger nucleases, transcription activator‐like effector nucleases, and the CRISPR‐associated (Cas) nuclease system 9 . Among them, CRISPR/Cas9 has become the top choice for gene editing due to its advantages of low cost, easy manipulation, and high efficiency.…”

Utilization of Genome Editing for Livestock Resilience in Changing Environment

“…The technique relies on a protein called Cas9, which acts like a pair of molecular scissors that can cut DNA at a specific location. Scientists can then insert, delete, or replace the cut DNA sequence with a desired sequence (Mehra and Kumar, 2022;Rasheed et al, 2022;Khiabani et al, 2023;Li et al, 2023), (b) Zinc Finger Nucleases (ZFNs): Zinc Finger Nucleases are engineered proteins that can bind to specific DNA sequences and cut the DNA at that location. Like CRISPR-Cas9, ZFNs can be used to add, delete or modify genes (Mehra and Kumar, 2022;Wani et al, 2023a), (c) TALENs: Transcription Activator-Like Effector Nucleases (TALENs) are another type of engineered proteins that can bind to specific DNA sequences and cut the DNA at that location, and TALENs work by using a DNA-binding domain that can be programmed to recognize a specific DNA sequence (Mehra and Kumar, 2022;Kumar and Kues, 2023;Wani et al, 2023a), (d) Homologous Recombination: Homologous Recombination is a technique that uses a DNA template to repair a broken DNA strand.…”

CRISPR gene technology—the next best thing in medicine

“…Using adult intestinal stem cells derived from CF patients [13] , one of the common CF mutations in intestinal organoids was effectively corrected. The CF transmembrane conductor receptor function was ultimately recovered after the mutation was corrected [14] .…”