gene editing

Phytomelatonin-mediated epigenetic and RNA regulatory networks in plant abiotic stress resilience.

This work proposes that phytomelatonin functions as an epigenetic and epitranscriptomic trigger capable of converting transient stress perception into durable transcriptional competence and outlines how single-cell multi-omics, targeted epigenome editing, epitranscriptomic profiling and field-scale validation of priming strategies can transform this conceptual framework into testable mechanisms and crop-improvement strategies.

The emerging roles of non-coding RNAs in cancer.

Non-coding RNAs (ncRNAs) regulate gene expression through transcriptional, post-transcriptional, and epigenetic mechanisms, shaping hallmarks of cancer, including metastasis, therapy resistance, and relapse. Carcinogenesis arises when aberrant ncRNA networks initiate malignant transformation and sustain oncogenic changes through epigenetic modifications, shifts in cell identity, failures in genome protection, metabolic changes, and alterations in the tumour microenvironment. Environmental exposures, combined with chronic inflammation, reorganise these networks early on, leading to the formation of premalignant fields and persistent epigenetic changes. The four major ncRNA classes, microRNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and PIWI-interacting RNAs (piRNAs) function as either oncogenes or tumour suppressors depending on the specific cancer type. Their stability, cell-type-specific expression, and presence in biofluids make them suitable candidates for biomarker discovery and liquid biopsy applications. Therapeutic strategies now include antisense oligonucleotides, small interfering RNAs, synthetic miRNA mimics, RNA aptamers, and aptamer-siRNA conjugates, which can either inhibit oncogenic ncRNAs or restore tumour-suppressive regulatory networks. CRISPR-based ncRNA modulation, including Cas9-mediated locus editing, CRISPR interference/activation, and Cas13-mediated transcript targeting, remains largely investigational because delivery, off-target activity, and an incomplete understanding of ncRNA context dependence continue to limit translation. High-throughput sequencing, single-cell transcriptomics, and computational modelling have accelerated the identification of cancer-related ncRNAs and elucidated their biological functions. This review examines how different types of ncRNAs contribute to cancer initiation, progression, and treatment resistance, and assesses their potential as diagnostic markers, prognostic factors, and therapeutic targets.

Functional assessment of inherited myeloid neoplasm-associated SAMD9L germline variants via Monoallelic CRISPR modelling.

While the majority of myeloid neoplasms are sporadic, the increasing application of germline genetic testing has led the World Health Organization to designate 'Myeloid malignancies with germline predisposition' as a distinct clinical entity, carrying major implications for clinical care and research. Germline variants in the Sterile alpha motif domain-containing 9-like (SAMD9L) gene are specifically associated with an increased risk for these malignancies. In this study, we functionally modelled two novel SAMD9L variants-p.N697Y and p.K1294*-alongside two previously reported variants (p.T233N and p.H880Q). We generated heterozygous knock-in cellular models in the Human leukemia (HL-60 myeloid cell line) for each variant using homology-directed repair-based Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR/Cas9) gene editing. Functional assays, focused on proliferation and protein translation, confirmed that the p.T233N, p.N697Y and p.K1294* variants all caused a decreased rate of protein translation. These results provide functional evidence to fine-tune the classification of these SAMD9L variants and significantly advance our understanding of the molecular mechanisms by which SAMD9L variants drive inherited myeloid neoplasms.

Targeting Cas9 to Perform Rescue Experiments in Stably Transduced Knockout HEK293 Cells.

CRISPR-Cas9 gene editing technology has revolutionized molecular biology. Often, this technology is employed to delete a gene encoding a protein of interest. The resulting phenotype provides valuable insight into the protein's function. The functional importance of the target protein can be confirmed by reintroducing the protein to restore the lost function (rescue). This is typically accomplished by introducing the protein-coding cDNA in trans using an expression vector. However, in knockout cell lines that stably express the CRISPR-Cas9 system, the newly introduced expression plasmid may also be cleaved by Cas9. The protocol presented here provides a strategy to circumvent this potential barrier to rescue experiments. This approach is demonstrated using HEK293 cells in which the gene encoding the E3 ubiquitin ligase scaffold protein CUL4B was disrupted by CRISPR-Cas9. Transduction of these cells with a guide RNA (gRNA) targeting the integrated Cas9 transgene resulted in the loss of detectable Cas9 protein. Cas9 ablation enabled restoration of CUL4B expression and function following introduction of a CUL4B expression plasmid. These results provide proof of concept for a broadly applicable approach to studying protein function through rescue experiments.

Advancing variant-agnostic liver-directed gene editing using homology-independent targeted integration.

A PepFect14 analog improves non‐viral CRISPR delivery in primary human cells to facilitate genome editing and repair

A PepFect14 analog is employed to deliver high-fidelity Cas9-ribonucleoproteins and non-viral repair templates into primary human skin cells to mediate gene editing and repair targeting genes underlying the group of genetic skin blistering disorders epidermolysis bullosa (EB).

Gene knockout of P450s CYP392A16 and CYP392D2 reveals unexpected in vivo effects and transcriptional rewiring in the extreme generalist Tetranychus urticae.

It is demonstrated that even resistance-associated detoxification genes that are considered functionally validated in vitro, can have strong context-dependent phenotypic effects in vivo, underscoring the complexity of resistance gene networks and highlighting the need to interpret CRISPR/Cas9 knockout phenotypes within the broader genetic and regulatory context.

Durability of CRISPR-Cas9 Gene Editing Targeting ANGPTL3 with CTX310.

In a phase 1a trial, one-time CRISPR-Cas9 gene editing with CTX310 was associated with no treatment-related dose-limiting toxic effects and with sustained reductions in ANGPTL3 and atherogenic lipids at 1 year.

Pan-Genomic Dissection of GH1 β-Glucosidases in Brassica rapa Identifies BrBGLU10 as an Important Regulator of Pollen Development

Findings establish BrBGLU10 as an important regulator of pollen development and a potential target for fertility-related applications via gene editing in B. rapa and related Brassica crops.

Non-invasive Ultra-early in Utero Detection and Precision CRISPR-mediated Correction of Monogenic Embryonic Mutations: A Critical Appraisal of a Hypothetical Therapeutic Framework

Whether that proposition that a pathogenic single-gene variant might be identified non-invasively at the earliest stage of pregnancy and corrected in situ before irreversible pathology develops is presently coherent as a therapeutic framework is examined.

Gene and nucleic acid therapies for MMUT-deficient methylmalonic acidemia: from AAV-mediated gene addition to LNP-mediated mRNA delivery and precision genome editing

Current therapeutic evidence is concentrated primarily in MMUT- deficient isolated MMA, and further progress will require safer and more durable delivery platforms, improved tissue targeting, robust long-term safety assessment, clinically meaningful endpoints, and careful evaluation of accessibility and cost.

Chinese geneticists across the Pacific: a century of scientific legacies in genetics and genomics

This historical review presents the breakthrough discoveries by prominent Chinese geneticists during the early pioneering generation, the molecular genetics era, the transition to human and medical genetics, and current progress in genetic and genomic medicine.