Comprehensive strategies for constructing efficient CRISPR/Cas based cancer therapy: Target gene selection, sgRNA optimization, delivery methods and evaluation

• Published in: Advances in colloid and interface science Vol. 341; p. 103497
• Main Authors: Paranthaman, Sathishbabu, Uthaiah, Chinnappa A., Md, Shadab, Alkreathy, Huda Mohammed
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2025
• Subjects: Animals; Cancer therapy; Cell-penetrating peptides; CRISPR-Cas Systems - genetics; CRISPR/Cas genome-editing; Extracellular vesicles; Gene Editing - methods; Genetic Therapy - methods; Humans; Neoplasms - genetics; Neoplasms - therapy; Non-viral vesicle; RNA, Guide, CRISPR-Cas Systems - genetics; Viral vesicle; PNPs; LNPs; Extracellular vesicles; CRISPR/Cas genome-editing; Viral vesicle; gRNA; CPPs; OSSC; TALEN; HDR; crRNA; CRISPR; ZFN; AAVs; EVs; IMNPs; TKIs; LVs; dCas9; Non-viral vesicle; Cas; sgRNA; Cancer therapy; Cas9; NHEJ; Cell-penetrating peptides; PAM; AdVs
• ISSN: 0001-8686; 1873-3727; 1873-3727
• DOI: 10.1016/j.cis.2025.103497

Cancer is a complicated disease that results from the interplay between specific changes in cellular genetics and diverse microenvironments. The application of high-performance and customizable clustered regularly interspaced palindromic repeats/associated protein (CRISPR/Cas) nuclease systems has significantly enhanced genome editing for accurate cancer modeling and facilitated simultaneous genetic modification for cancer therapy and mutation identification. Achieving an effective CRISPR/Cas platform for cancer treatment depends on the identification, selection, and optimization of specific mutated genes in targeted cancer tissues. However, overcoming the off-target effects, specificity, and immunogenicity are additional challenges that must be addressed while developing a gene editing system for cancer therapy. From this perspective, we briefly covered the pipeline of CRISPR/Cas cancer therapy, identified target genes to optimize gRNAs and sgRNAs, and explored alternative delivery modalities, including viral, non-viral, and extracellular vesicles. In addition, the list of patents and current clinical trials related to this unique cancer therapy method is discussed. In summary, we have discussed comprehensive start-to-end pipeline strategies for CRISPR/Cas development to advance the precision, effectiveness, and safety of clinical applications for cancer therapy. [Display omitted] •This review presents a comprehensive framework to enhance CRISPR/Cas-based cancer therapies, tackling key challenges.•It details strategies for target gene identification, gRNA/sgRNA optimization, and various delivery methods.•Concludes with insights into patents and clinical trials, showcasing CRISPR/Cas's future potential in cancer therapy.