2026· Methods in molecular biology· Vol 3005, pp.
383-401
· 0 citations
Medicine
TL;DR
An overview of the CRISPR screen methodology, which involves introducing guide RNAs targeting specific genes into cells, followed by phenotype selection and analysis, is presented, providing valuable insights into gene function and potential therapeutic targets.
Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide, characterized by genetic heterogeneity and the accumulation of mutations in key oncogenes and tumor suppressor genes. CRISPR-Cas9 technology has greatly advanced genetic research by enabling precise genome editing. This review focuses on the innovative applications of CRISPR-Cas9 in CRC research, particularly its role in identifying novel therapeutic targets, elucidating mechanisms of drug resistance, and uncovering metabolic and stem cell pathway alterations in tumorigenesis. We highlight the diverse CRISPR systems, including Cas9, Cas12, Cas13, and advanced variants such as CRISPR activation (CRISPRa), CRISPR interference (CRISPRi), base editing, and prime editing, which have expanded gene knockout studies and enhanced our understanding of CRC. Despite these breakthroughs, challenges such as off-target effects and delivery limitations remain. Ongoing efforts to refine CRISPR technology aim to enhance its precision and clinical applicability, ultimately paving the way for more effective and personalized treatment strategies for CRC. In this review, we explore these advances and focus on the latest developments in CRISPR-based approaches for CRC treatment.
Mohadeseh Hassannia, P. Amirifar· Cancer Treatment and Researc...· 0 citations
Key methodological steps for achieving high-efficiency lentiviral transduction and selection are described, enabling the successful application of EPIKOL CRISPR screens in chemoresistant TNBC models.
O. Yedier-Bayram, Elif Ayca Guvener, T. Bagci-Onder· Journal of Visualized Experi...· 0 citations
How CRISPR-enabled functional genomics approaches can reveal unexpected cancer dependencies and resistance mechanisms is outlined, and emerging applications of CRISPR-based diagnostics in oncology that convert precise nucleic acid sequence recognition into rapid mutation detection are discussed.
S. Grigg, Carolyn Shembrey, M. Fareh et al.· Nature Reviews Clinical Onco...· 1 citation
As the field of anticancer drug development is constantly changing, combining synthetic lethality with epigenetic modifiers opens up new possibilities for targets that fall outside the traditional drug target range. Enzymes involved in DNA methylation and histone modification are examples of epigenetic modifiers that promote gene expression without altering the DNA sequence of the gene. These mechanisms play a key role in carcinogenesis when they are altered, as they downregulate tumor suppressor genes and overexpress oncogenic pathways. Synthetic lethality is a phenomenon in which simultaneous mutations or perturbations of two genes result in cell death, but alterations to one gene alone do not cause cell death. It was first observed in genetic research conducted on model organisms, such as fruit flies and fungi. The most well-known example of this idea from the perspective of cancer treatment is PARP inhibitors, which are effective in tumors with BRCA1/2 mutations, where further failure of DNA repair results in cell sensitization. Building on the concept of synthetic lethality, current research focuses on exploiting epigenetic flaws that are common in cancer cells. For example, when chromatin remodelers or methyltransferases cease to function, malignant cells undergo genetic rewiring, rendering them vulnerable to treatment. Recent research has produced some striking examples of synthetic-lethal drug interactions and biomarkers used in metagenomics for personalized medicine by targeting the secondary pathways used by cancer cells as a result of primary loss-of-function mutations, which selectively kill cancer cells while sparing healthy cells. The discovery of actionable epigenetic dependencies and overcoming tumor heterogeneity remain the largest challenges in translating these fascinating scientific discoveries to the clinic. The convergence of epigenetic modulators with synthetic-lethality-based therapeutic architectures is poised to define a transformative paradigm in precision oncology. By orchestrating multilayered perturbations across chromatin-regulatory networks, DNA damage-response pathways, and context-specific vulnerability nodes, this integrative strategy surpasses the limitations of conventional target-centric pharmacology and enables mechanistically rational, synergistic antitumor interventions.
Venkatesh Kamath, Vasudev Pai, Bhavana Bhat et al.· Current Topics in Medicinal...· 0 citations
Overall, CRISPR/Cas9 represents a promising yet evolving platform in oncology, with its future clinical success dependent on achieving a balance between precision, safety, scalability, and long-term therapeutic durability.
C. Ng, Sakina Mustafa, X. Y. Yap et al.· Frontiers in Oncology· 0 citations
CRISPR has emerged as a next-generation gene-editing tool with the potential to target the molecular pathways associated with ageing and related disorders. It functions through RNA-guided Cas nucleases, directing DNA cleavage and utilizing the native DNA repair machinery for genetic manipulations. Advances in CRISPR technology have significantly enhanced the precision and flexibility of techniques for genome editing. The enzyme Cas9's ability to cut DNA at exact site has revolutionized genome editing by enabling accurate modifications within living eukaryotic cells. This review critically examines recent developments in CRISPR-based technologies, including Cas9, Cas12, base editing, prime editing, and CRISPR-mediated gene regulation. It highlights their rising applications in ageing research, with more emphasis on neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. The review also discusses the major pharmacological and translational challenges that currently limit clinical applications, including inefficient tissue-specific delivery, off-target genome editing, immunogenicity, manufacturing complexity, and long-term safety concerns. Also, recent progress in both, viral and non-viral delivery methods are critically evaluated, including adeno-associated viruses, lentivirus vectors, lipid nanoparticles, gold nanoparticles, exosomes, electroporation, and microinjection, is thoroughly discussed to highlight their therapeutic potential and translational limitations. Current studies indicate that CRISPR-based approaches have preclinical potential for targeting important hallmarks of ageing, particularly genomic instability, telomere attrition, and mitochondrial dysfunction. Other hallmarks of ageing, such as stem cell exhaustion, epigenetic modifications, and microbiome changes, are at earlier stages of development. Overall, this review describes future strategies for developing safe, precise, and clinically translatable CRISPR-based treatments to promote healthy ageing.
Sakshi Rathore, Akash Gupta, Kamal Shah et al.· Ageing Research Reviews· 0 citations