Colorectal cancer (CRC) continues to be one of the most prevalent gastrointestinal malignancies on a global scale. Despite advancements in targeted therapy and immunotherapy that have enhanced outcomes for certain patients, primary and acquired drug resistance remain significant challenges that impede clinical efficacy. Metabolic reprogramming is widely acknowledged as a fundamental hallmark of cancer, facilitating survival advantages for tumor cells by restructuring glucose, amino acid, and lipid metabolic networks. Critically, this reprogramming directly fosters a drug-tolerant phenotype by modifying the flux of metabolic intermediates. Simultaneously, aberrant epigenetic modifications—including the remodeling of DNA methylation, altered histone acylation, and dysregulated non-coding RNA networks—play an indispensable role in the establishment and maintenance of resistance to targeted therapies in CRC. Notably, metabolic reprogramming and epigenetic remodeling do not function independently. Recent research indicates that cellular metabolites serve as substrates or cofactors for the regulation of epigenetic modifying enzymes. Conversely, alterations in the epigenome feedback to regulate the transcription of metabolic enzymes. This interplay establishes a bidirectional regulatory mechanism.
Accelerating climate change is driving unprecedented increases in the frequency and severity of abiotic stresses, threatening global food security and demanding new strategies for crop resilience. Plants have evolved a sophisticated molecular response to recurrent stress through epigenetic priming, where prior exposure induces chromatin-level alterations encoded through DNA methylation, histone modifications, and small RNA pathways. These imprints can persist somatically across cell divisions and, in some cases, be transmitted to subsequent generations. CRISPR/dCas9-based epigenome editing provides the means to install, modify, or erase these same marks at defined loci without altering the underlying sequence. This mini review provides an evidence-based survey of plant dCas9 epigenome editing, distinguishing what each effector class has actually been demonstrated to achieve in planta from what has been shown only in mammalian systems or remains inferred. I map the translational barriers between Arabidopsis proof-of-concept and crop deployment, survey what has been achieved in rice, maize, and tomato, organize the toolkit around the distinct memory logics (type I persistent versus type II poised) it must serve, and identify the priorities that would advance the field from demonstrated single-locus edits toward heritable, deployable modifications. By separating demonstrated capability from extrapolation, this synthesis provides a realistic basis for applying epigenome editing to sustainable agriculture and climate-resilient crop production.
Sugar crops including sugarcane, sugar beets, and sweet sorghum, are fundamental for global sucrose supply and crucial feedstocks for bioethanol production and industrial bioproducts. However, yield and sucrose accumulation are significantly influenced by abiotic stresses, including drought, salinity/alkalinity, heat, chilling/freezing, and heavy metal toxicity. The polygenic complexity of stress tolerance, particularly in sugarcane with its large, polyploid, and haplotype-diverse genome, presents significant challenges for conventional breeding, rendering it slow and imprecise. This review combines advances in genome-editing technologies, including CRISPR/Cas systems (Cas9 and Cas12a), along with transcriptional techniques (CRISPRa/i), base editors, and prime editors. These tools, combined with the progress in reference genomes, haplotype-resolved assemblies, and pangenomes, have facilitated precise gene discovery, genomic selection, and targeted editing. CRISPR-based approaches offer promising avenues for enhancing key stress-responsive pathways governing oxidative homeostasis, membrane integrity, osmotic regulation, hormone signaling, and ion-homeostasis in sugar crops. Prominent candidate targets include transcription factors (TF), signaling components, raffinose biosynthesis genes implicated in freezing tolerance, regulators of salt and alkaline stress, and determinants of heavy metal tolerance and sequestration. Translational challenges, such as off-target effects, low transformation efficiencies, and editing multiple homologues or haplotypes, are being addressed such as DNA-free ribonucleoprotein (RNP) delivery, high-throughput screening, haplotype-specific target design, and haplotype-aware guide RNA engineering based on unique genetic variation sites across different gene haplotypes. Collectively, the integration of advanced CRISPR toolkits with enhanced genomic information resources and phenotyping capabilities holds significant potential for accelerating the development of precision-bred, multi-stress-resilient sugar crops that sustain yield, quality, and industrial value under abiotic stress conditions.
Chimeric antigen receptor (CAR)-based immunotherapy has transformed cancer treatment by enabling engineered immune cells to selectively recognize and eliminate malignant targets, with CAR-T cells establishing the first major clinical success in this field. However, severe associated toxicities, logistical complexities of autologous manufacturing, and high rates of antigen-loss relapse have accelerated the development of alternative effector platforms such as CAR-NK cells. CAR-NK cells combine antigen-directed cytotoxicity with the innate antitumor functions of natural killer cells, offering a potentially safer and more readily available off-the-shelf therapeutic strategy. Nevertheless, the clinical efficacy of CAR-NK therapy is constrained by key biological and translational hurdles: brief in vivo persistence, inadequate trafficking and solid tumor infiltration, severe susceptibility to tumor microenvironment-mediated immunosuppression, and antigen escape driven by tumor heterogeneity. These biological constraints are compounded by technical bottlenecks, including cell source variability, limited large-scale expansion capacity, ex vivo functional exhaustion, and an absence of standardized manufacturing protocols. To address these multi-faceted barriers, recent advances in genome engineering, systems biology, and computational modeling provide promising routes forward. Given that prolonged exposure to immunosuppressive niches induces functional impairment, genome-editing strategies targeting inhibitory checkpoints and persistence programs are crucial; however, enhancing persistence requires stringent safeguards to mitigate toxicity risks from constitutive signaling or genomic instability. Thus, engineering next-generation CAR-NK cells necessitates balancing extended longevity, potent cytotoxicity, and safety. Multiomics approaches facilitate deeper characterization of CAR-NK functional states and tumor–immune interactions, whereas artificial intelligence–guided engineering integrates high-dimensional biological and manufacturing data to optimize receptor design, predict therapeutic performance, and accelerate translational workflows. Collectively, these innovations support a paradigm shift toward programmable CAR-NK cells that can dynamically sense, adapt, and respond to tumor evolution, driving the transition toward autonomous, highly scalable, and precisely controllable cellular immunotherapies in precision oncology.
Given the growing emphasis on the efficient application of CRISPR/Cas9 in plant genome editing, there is a need for methodologies that enhance targeted mutagenesis efficiency. In the present study, we utilized stable transgenic maize callus tissues harboring active CRISPR/Cas9 vector systems targeting either the abnormal cytokinin response 1 (ARE1) gene or the potassium channel gene (KAT2). The presented DNA sequencing data indicate that the KAT2-targeting vector, which utilizes a 3′-end truncated guide RNA, retained mutagenic activity, albeit at a notably low frequency. We quantified mutagenic events following exposure to elevated temperatures (37 °C for 48 h) and various chromatin-modifying compounds, including sodium butyrate (NaBt), nicotinamide (NA), and 5-azacytidine (AZA). High-temperature treatment alone increased the frequency of nucleotide deletions (5.8-fold relative to the untreated samples) in the ARE1 target gene. Conversely, in the KAT2 gene, both deletions and insertions were stimulated (6.4 and 6.2-fold, respectively). As a novel approach, we evaluated the mutagenic efficacy of chromatin-modifying compounds administered in conjunction with heat treatment. The combined application of NaBt and AZA with elevated temperature was particularly effective in increasing insertion frequencies; in the ARE1 (with NaBt + heat: 17.3-fold) and KAT2 (with AZA + heat: 21.4-fold) genes. Cell cycle analysis of suspension-cultured cells revealed that chromatin-modifying treatments altered cell cycle progression, evidenced by an accumulation of cells in the G2 phase and a significant reduction in the number of mitotic cells. Subsequent DNA sequencing demonstrated a differential increase in the induction of deletions and insertions, alongside an expansion of the deleted regions.
