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.
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
Cancer immunotherapy has shown significant promise in certain patient populations, but further advancements are needed to extend its benefits to a wider range of patients. Clustered regularly interspaced short palindromic repeats (CRISPR)-based editing has rapidly evolved in recent years, enabling its transition into direct therapeutic applications. This review summarizes recent progress in applying CRISPR systems in vivo for cancer immunotherapy, focusing on approaches that target cancer cells and the tumor microenvironment, as well as those that directly engineer immune cell populations themselves. Novel CRISPR editing platforms and strategies enabling multiplexed editing have also recently demonstrated promising impacts on driving antitumor immunity, however, the platforms investigated are still in the early stages and further investigation will be needed to robustly assess the potential for clinical translation. Future work can expand the array of therapeutic targets by incorporating data from functional genomics and must also carefully evaluate both editing modalities and delivery systems to optimize efficacy, safety, and scalability.
Cole W. Christopher, Xiaoyu Zhou· Frontiers in Immunology· 0 citations
Immuno-oncology has reshaped the therapeutic landscape of cancer treatment by shifting focus from directly targeting tumor cells to mobilizing the immune system against malignancies. Among the most transformative advances in this field is the development of chimeric antigen receptor T-cell therapy, which has demonstrated remarkable efficacy in hematologic cancers. However, persistent challenges such as limited durability, immune escape, toxicity, and poor performance in solid tumors have constrained its broader clinical impact. The emergence of clustered regularly interspaced short palindromic repeats (CRISPR) genome editing has introduced a powerful and versatile platform for engineering immune cells with enhanced specificity, persistence, and functionality. CRISPR-based approaches enable precise gene knockout, targeted gene insertion, epigenetic modulation, and multiplex editing, allowing researchers to redesign immune cells at multiple regulatory levels. These capabilities have significantly advanced CAR-T cell engineering and have catalyzed the development of next-generation immune effectors, including natural killer cells, macrophages, and stem cell-derived immune populations. Furthermore, CRISPR technology has opened new avenues for overcoming the immunosuppressive tumor microenvironment, improving safety profiles, and enabling scalable, off-the-shelf therapies. This review provides a comprehensive examination of CRISPR applications in immuno-oncology, with an emphasis on CAR-T optimization and the engineering of next-generation immune cells. It discusses mechanistic foundations, technological innovations, preclinical and clinical advancements, safety considerations, and future directions. Collectively, CRISPR-driven immune engineering represents a paradigm shift toward more precise, effective, and accessible cancer immunotherapies.
Adewale Adeleke· International Journal for Sc...· 0 citations
Cancer research has undergone a transformative change with the advent of high-throughput genomic technologies. Advances in next-generation sequencing accelerated the identification of somatic and germline alterations that drive tumorigenesis enabling the transition from traditional histology-based cancer classification to molecularly informed precision oncology. Large-scale sequencing initiatives and clinical genomic profiling facilitated the development of companion diagnostic assays and targeted therapies. Beyond targeted therapies, genomic innovations have also catalyzed the emergence of novel therapeutic strategies including immunogenomics-driven immunotherapies, RNA-based therapeutics, cancer vaccines and genome editing technologies based on CRISPR-Cas systems. This review summarizes the major technological developments in cancer genomics, including sequencing platforms, transcriptomic profiling, liquid biopsy, and functional genomic screening, and highlights the utility of these innovations in discovery of actionable biomarkers and next-generation therapeutic strategies. Collectively, these advances underscore the central role of genomic technologies in driving the evolution of precision oncology toward more personalized and effective cancer treatment strategies.
Hanumappa Ananda, Sadhu R. Sahana, Shruthi R. Murthy et al.· Frontiers in Genetics· 0 citations
A comprehensive review of delivery modalities of CRISPR systems solely in vivo that underpin their therapeutic translation and outlines the remaining barriers to durable, tissue-selective, and broadly deployable CRISPR therapeutics is provided.
L. Martin, Jure Bohinc, Alessandra Recchia et al.· International Journal on Bio...· 0 citations
Background: Leukemia is a cancer of hematopoietic stem cells in the bone marrow. It is classified as lymphoid or myeloid, and as acute (rapid onset) or chronic (slow progression). Advances in CRISPR technology enable deeper study of leukemia biology and therapeutic targets. While AML, CML, ALL, and CLL are distinct, comparative studies highlight shared and unique features. Identifying key pathways across subtypes may yield improved treatments. Our lab has established a unique CRISPR Activation (CRISPRa) platform for gain-of-function screening applications. We hypothesize that genome-wide CRISPRa screening will uncover subtype-specific genetic dependencies and drug resistance mechanisms, offering novel therapeutic insights.
We engineered leukemia cell lines by nucleofecting a self-selecting CRISPRa PiggyBac plasmid carrying a blasticidin resistance gene, enabling selection of CRISPRa-competent cells. Functionality was validated using lentiviral delivery of sgRNAs targeting cell surface markers. Cells were transduced with our whole-genome CRISPRa library, Sonata, at MOI 0.4, followed by puromycin selection to enrich for sgRNA-expressing cells. Post-transduction, cells were harvested at various timepoints for sequencing to track sgRNA abundance and identify growth-modulating genes.
We engineered and validated four CRISPRa-competent leukemia lines: K562 (CML), Jurkat (T-ALL), THP-1 (AML), and HL60 (AML). We have completed whole-genome screening campaigns in the CML and T-ALL backgrounds, identifying hundreds of shared and context specific growth modifiers. Functional validation, pathway analysis, and potential clinical significance is ongoing, as is expansion of our screens with the AML contexts.
Conclusions: Our screens reveal genes that, upon activation, influence leukemia cell growth. These findings support discovery of new therapeutic targets and enhance understanding of leukemia subtype biology, guiding future personalized treatment strategies.
The Cole Foundation, FHMR
Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)
Jeffrey Sullivan, Xiaozen Wen, Gabriela Flores-Vargas et al.· Journal of Immunology· 0 citations