gene editing

Generation of two homozygous iPSC lines carrying variants of uncertain significance in LMNA associated with cardiomyopathy.

Variants of uncertain significance (VUS) in the LMNA gene represent a major challenge in clinical genetics, as insufficient functional evidence limits their interpretation and clinical decision-making in laminopathies, including dilated cardiomyopathy (DCM). Here, we generated two isogenic induced pluripotent stem cell (iPSC) lines carrying homozygous LMNA variants, c.293A > G (p.Glu98Gly) and c.439G > A (p.Ala147Thr) by prime editing of a healthy donor iPSC line. Both variants are located within Coil 1B domain of lamin A. The edited iPSC lines retain normal morphology, pluripotency, genomic integrity, and trilineage differentiation capacity, providing a valuable platform for functional characterization and potential clinical reclassification of LMNA VUS.

Design of base editing systems with tunable editing hotspots within a defined window through sgRNA engineering.

Base editors (BEs) enable efficient A-to-G or C-to-T conversions without double-stranded DNA cleavage, but their editing windows remain difficult to tune, limiting genome engineering flexibility. Here, we engineered CRISPR/Cas12b sgRNA by introducing MS2 hairpins to recruit an MS2-N55K-cytidine deaminase-UGI complex, enabling programmable control of the editing window. Three modified sgRNAs were generated by replacing two loop regions, each producing distinct editing hotspots in E. coli. The AID*Δ-MSBE system (sgRNA1.1) generated a window near the PAM with peak activity at C7-C9, while the CDA-MSBE system (sgRNA1.2) produced a distal window with peak activity at C20-C23. Both systems exhibited identical editing patterns in Bacillus subtilis. A dual-orthogonal system (MS2 and PP7) was constructed to simultaneously recruit two deaminase complexes, restoring the classic dCas12b CBE editing pattern. Rifampicin resistance assays confirmed high targeting specificity with low off-target effects. As proof of concept, the MSBEs were successfully employed for the flexible reprogramming of sfGFP fluorescence and the targeted evolution of the endogenous gene rpsE, respectively. Collectively, we developed the MSBEs with tunable editing hotspots, providing innovative tools to enhance the flexibility and accessibility of BEs for genome engineering.

Engineering HSV-1 for Gene Therapy, Oncolytic Immunotherapy, and Neuroscience.

Herpes simplex virus type 1 (HSV-1) has emerged as a versatile platform for gene delivery, oncolytic immunotherapy, and neural circuit mapping. Its large genome, broad tropism, and engineering flexibility enable delivery of large or multi-component payloads that exceed the capacity of many conventional viral vectors. HSV-1-based vectors span a continuum of architectures, each representing a distinct design space shaped by trade-offs among replication competence, payload size, immune engagement, biosafety, and manufacturing robustness. Advances in bacterial artificial chromosome recombineering, CRISPR-based editing, and synthetic genome assembly, together with insights from structural and systemic biology, have accelerated the transition from empirical vector construction to more rational programmable genome design. These technologies enable modular control of viral entry, transcription, genome replication/maintenance, and host immune interactions. Clinical successes such as T-VEC, G47Δ, and B-VEC have validated the clinical potential of HSV-1 engineering, yet broader translation remains limited by antiviral immunity, inefficient delivery, epigenetic silencing, genome instability, and manufacturing challenges. In this review, we illustrate how HSV-1 has evolved from a naturally neurotropic virus into a versatile biomedical tool, whose therapeutic and research potential emerges from the precise matching of viral properties with disease-specific requirements, delivery contexts, and functional objectives.

Can host genetics transform the sustainable control of tropical theileriosis? Insights from the Tick-Theileria interface.

Tropical theileriosis, caused by the tick-transmitted apicomplexan parasite Theileria annulata, remains a major constraint on cattle production across North Africa, the Mediterranean basin, the Middle East and South Asia. Current control depends on acaricides, the theilericidal drug buparvaquone and live attenuated schizont vaccines, but acaricide resistance, buparvaquone-resistance mutations and the logistical demands of vaccination are eroding the sustainability of these tools. Host genetics offers a complementary and durable alternative. Indigenous Bos indicus breeds are consistently more resistant to ticks and tolerate T. annulata infection better than exotic Bos taurus cattle, and this advantage has a measurable heritable component. Unlike previous reviews, which treat tick resistance, T. annulata immunobiology and livestock genomic selection as separate subjects, we integrate all three and assess host genetics specifically against the failure modes of current control. We review the tick, parasite and host interface, the evidence for natural resistance, and the genetic and immunological mechanisms involved, including signal-regulatory protein, bovine major histocompatibility complex class II and inflammatory pathway genes. We then assess whether genomic selection, multi-omics, machine learning and gene editing can translate these mechanisms into resistant cattle, and we weigh the biological, economic and infrastructural barriers to implementation. The evidence indicates that host genetics will not replace existing control but could reduce reliance on acaricides and chemotherapy. That contribution remains prospective rather than demonstrated: no resistance marker for T. annulata has yet been validated, prediction accuracies are moderate and transfer poorly between breeds, and no endemic production system has implemented selection for resistance.

Gene editing of alveolar organoids reveals AT2 dysfunction due to endosomal SFTPC accumulation.

Endosomal dysfunction and polarity loss are established as drivers of SFTPC-I73T-mediated epithelial injury and mechanisms that may underlie AT2 dysfunction in disease more broadly are highlighted.

Global genetic modifiers of fetal hemoglobin in sickle cell disease: Systematic review and meta-analysis.

Evidence on genetic determinants of Hb F and quantified effects of key variants through meta-analysis is synthesized, supporting genotype-guided therapeutic, and genome-editing strategies.

CRISPR gene therapy: a review

Clinical applicability is limited by issues such off-target effects, PAM sequence restrictions, DNA damage-induced toxicity, and immunological responses to Cas proteins, despite its wide therapeutic potential, but improvements in delivery methods and high-fidelity Cas9 variations are being addressed.

Intranasal CRISPR lipid nanoparticles targeting MAPK9 attenuate neuroinflammation after traumatic brain injury

Traumatic brain injury (TBI) induces a sustained neuroinflammatory response involving activated microglia and infiltrating myeloid cells, contributing to secondary brain damage and long-term neurological dysfunction. Modulating these inflammatory responses toward a more reparative phenotype represents a promising therapeutic strategy, but achieving targeted delivery within the injured brain remains a major challenge. Here, we developed a targeted, non-viral gene-editing platform using lipid nanoparticles (LNPs) encapsulating CRISPR-Cas12a components directed against MAPK9, a key mediator of inflammatory signaling. LNPs were functionalized with an Iba-1 antibody to enhance targeting of Iba-1 + myeloid cells following intranasal administration. In primary bone marrow-derived macrophages and primary microglia, CRISPR-mediated MAPK9 targeting reduced MAPK9 expression and suppressed pro-inflammatory activation, decreasing iNOS, NLRP3, CD80, and CCL2 while increasing the anti-inflammatory/reparative markers CD206 and Arg1. In a mouse model of TBI, intranasally delivered Iba-1-targeted CRISPR-LNPs showed preferential association with Iba-1 + cells compared with NeuN+ neurons in the injured cortex and reduced MAPK9 expression within Iba-1 + cells. CRISPR-LNP treatment attenuated microglial/macrophage activation, reduced pro-inflammatory cytokine expression, and decreased iNOS+/Iba-1 + cells while increasing CD206+/Iba-1 + cells in the peri-contusional cortex, supporting a shift toward a less inflammatory phenotype. Treatment also exhibited a favorable safety profile, with no detectable toxicity in the major organs examined. Together, these findings demonstrate that intranasal delivery of Iba-1-targeted CRISPR-LNPs enables effective MAPK9 modulation in Iba-1 + myeloid cells within the injured brain and attenuates acute neuroinflammation following TBI. This non-invasive therapeutic platform provides a promising approach for targeted modulation of neuroinflammatory responses after brain injury.

Eimeria in chickens: Recent advances in host-pathogen interactions, molecular insights, and control strategies.

Avian coccidiosis, caused by Eimeria spp., remains a major parasitic disease of poultry and imposes significant economic burdens on the global poultry industry. This review systematically synthesizes key advances over the past decade concerning host-Eimeria interactions, molecular regulatory mechanisms, and novel control strategies, while contextualizing these findings with earlier seminal discoveries. In recent years, novel diagnostic tools based on molecular detection and antigen capture have emerged, offering improved sensitivity and interspecies specificity over conventional methods. These techniques complement traditional approaches relying on oocyst morphology and histopathology, and provide critical support for accurate assessment of field infection status, species and genotype discrimination, monitoring of drug-sensitivity shifts, and elucidation of transmission dynamics. Epidemiological investigations have further revealed the impacts of rearing management, environmental temperature and humidity, host genetic background, and gut microbiota composition on infection kinetics, underscoring the necessity of integrating biosecurity and precision management into regionally tailored control programs. Utilizing chicken embryo and chick infection models, in conjunction with CRISPR/Cas9 gene editing, single-cell transcriptomics, and high-resolution proteomics, researchers have gained deeper insights into key regulatory genes governing invasion, asexual multiplication, and gametogenesis, as well as invasion-related effector molecules and resistance-associated markers, thereby laying a foundation for the identification of novel intervention targets. In immunology, growing knowledge of the intestinal epithelial barrier response, Th1/Th17 polarization, regulatory T-cell function, and immune evasion strategies (e.g., antigenic variation and downregulation of host antigen presentation) provides a theoretical basis for the rational optimization of subunit vaccines and live oocyst vaccines. On the therapeutic front, novel combination regimens of conventional anticoccidials and plant-derived bioactive compounds have shown efficacy in reducing oocyst shedding and alleviating intestinal lesions, while nanoparticle-based targeted delivery systems and adjuvant combination strategies are being explored to enhance drug bioavailability or vaccine-induced protective immunity. Nevertheless, the effective integration of ever-expanding omics data, immune-protective mechanisms, and field-applicable control measures, while concurrently addressing drug residues and resistance management, remains a central challenge for achieving sustainable coccidiosis control.

Gene Editing Therapy Developer Scribe Therapeutics Closes $155.5M IPO.

Genome-wide Identification of CXE Gene Family in Soybean and Functional Characterization of GmCXE31 in Lipid Biosynthesis and Salt Tolerance.

This study elucidates the multifaceted roles of GmCXE31 in coordinating soybean salt tolerance, lipid metabolism and agronomic traits, providing theoretical and genetic resources for salt-tolerant and high-quality soybean molecular breeding.

Programmable Domestication: CRISPR, Pan‐Genomics and System Level Engineering for Next‐Generation Crops

ABSTRACT Global agriculture is increasingly challenged by climate instability, genetic erosion, emerging pathogens and rising food demands, exposing the limitations of conventional breeding and traditional domestication strategies. Recent advances in CRISPR‐based genome editing, pangenomic, synthetic biology, artificial intelligence (AI)‐assisted breeding and predictive phenomics are transforming de novo domestication from a slow evolutionary process into a programmable framework for rational crop redesign. This review synthesises recent advances in programmable de novo domestication and highlights how crop wild relatives and underutilised germplasm can be harnessed to develop resilient, climate‐adaptive and sustainable crop systems. The integration of multiplex genome editing, pan‐genomic variation discovery, AI‐driven genomic prediction and predictive breeding enables precise engineering of key domestication traits governing plant architecture, yield potential, stress resilience and nutritional quality. Furthermore, we propose a trajectory‐based framework for programmable domestication comprising Adaptive Rescue, Agronomic Refinement and Novel Chassis Engineering, which illustrates distinct evolutionary pathways, engineering complexity and crop redesign objectives. We also examine the major system level challenges that constrain programmable domestication, including cryptic genetic variation, epistasis, gene regulatory network complexity, genotype phenotype predictability, biodiversity conservation and regulatory considerations. Collectively, programmable domestication represents a transformative shift from conventional crop improvement towards system‐level engineering of next‐generation crops, providing a strategic foundation for enhancing global food security, agricultural sustainability and environmental resilience in the face of accelerating climate change.