Frontiers in genome editing最新文献

{"title":"R2Tg retrotransposon mediated targeted integration of exogenous genes into the 28S rDNA locus in mammalian cell lines for PKU gene therapy.","authors":"Xiuyuan Wang, Ziqi Wen, Rongjun Xu, Wenjing Shi, Yiling Long, Xingri Zhan, Rui Su, Jia Fei, Yumin Li","doi":"10.3389/fgeed.2026.1893815","DOIUrl":"10.3389/fgeed.2026.1893815","url":null,"abstract":"<p><p>Phenylketonuria (PKU) remains a challenging genetic disorder due to the absence of therapeutic strategies capable of achieving sustained restoration of phenylalanine hydroxylase (PAH) function. Here, we employed an R2 retrotransposon in <i>Taeniopygia guttata</i> (R2Tg)-based system to enable targeted integration of exogenous PAH mRNA into the 28S ribosomal DNA (rDNA) locus of the host genome, with the goal of rescuing PAH deficiency caused by loss-of-function mutations. Experimental results demonstrated that lipid nanoparticle (LNP) delivery of R2Tg facilitated efficient site-specific integration of EGFP and PAH transgenes in mammalian cells, significantly elevating PAH mRNA and protein expression without disrupting endogenous 28S rRNA levels. These findings support the potential of this strategy as a precise genomic modification tool with promise for lifelong correction of PAH deficiency.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1893815"},"PeriodicalIF":5.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621248/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149083129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancements in CRISPR-based <i>in vivo</i> gene therapy for hemophilia.","authors":"Xiaoyu Zhang, Kaidi Xu, Shihui Zou, Zhiyao Zhu, Shuzhi Teng, Ping Huang","doi":"10.3389/fgeed.2026.1924733","DOIUrl":"10.3389/fgeed.2026.1924733","url":null,"abstract":"<p><p>Hemophilia is an X-linked hereditary bleeding disorder caused by loss-of-function mutations in the genes encoding coagulation factors, leading to excessive bleeding and potentially being life-threatening. Currently, regular treatment for hemophilia is the infusion of recombinant blood coagulation factors. This approach is not only costly but can also give rise to complications such as the development of neutralizing antibodies (Nabs), which impede the therapeutic outcome. Hemophilia is a monogenic disease, making gene therapy a promising curative measure that includes gene addition and gene editing approaches. Adeno-associated virus (AAV) vectors have gained massive attention as the premier delivery vehicle for clinical gene therapy due to their diverse tissue tropisms dictated by the natural and engineered AAV capsids, their excellent safety profile since AAVs do not cause any known human diseases, low immunogenicity, and long-lasting gene expression. Several AAV-based gene addition therapies have been approved for hemophilia B and hemophilia A, and dozens of similar AAV-based clinical trials are underway. Gene editing technologies, like Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), have shown immense potential in the treatment of genetic diseases. CRISPR/Cas9-mediated transgene integration provides critical advantages over regular gene insertion therapy by enabling site-specific, targeted genomic integration rather than random integration. Clinical trials focusing on AAV-CRISPR/Cas9-mediated knock-in of human genes represent the frontier of <i>in vivo</i> genetic medicine, and one such prominent ongoing clinical trial for hemophilia B is designed to insert the human Factor IX gene into hepatocytes. This review outlines recent advances and feasible strategies of CRISPR/Cas9-based genome editing for hemophilia therapy, highlights the potential applications of next-generation gene editing and vector delivery technologies, and offers new insights for the advantages, limitations, and future directions for these novel treatment modalities.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1924733"},"PeriodicalIF":5.9,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13601842/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149042400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical translation of CRISPR-Cas9 therapeutics in cancer and inherited genetic disorders.","authors":"Khushi Bashir, Prathiksha Vasudev, Vikas Chhetri, Devika Prasad Dubhashi, Yash Gurunath Teli, Rhitam Biswas, Sudha Ramaiah, Anand Anbarasu","doi":"10.3389/fgeed.2026.1868779","DOIUrl":"10.3389/fgeed.2026.1868779","url":null,"abstract":"<p><p>CRISPR-Cas9, adapted from the bacterial Type II CRISPR adaptive immune system, functions as a programmable RNA-guided endonuclease that employs a single-guide RNA to direct Cas9 to specific genomic loci. CRISPR-Cas9 has transformed targeted genome editing by replacing complex protein engineering with programmable Watson-Crick base pairing between the guide RNA and target DNA. This review was developed following a structured literature search of major biomedical databases and clinical trial registries to synthesize current evidence on the therapeutic applications of CRISPR-Cas9 in oncology and inherited genetic disorders. Clinical studies of <i>ex vivo BCL11A</i>-enhancer editing have shown fetal hemoglobin reactivation, with most evaluable participants with sickle cell disease remaining free of severe vaso-occlusive crises for the prespecified period and most evaluable participants with transfusion-dependent β-thalassemia achieving sustained transfusion independence. <i>In vivo</i> reductions in circulating transthyretin protein levels have been achieved for transthyretin amyloidosis via lipid nanoparticle delivery, while clinically meaningful improvements in selected measures of visual function were observed in a subset of patients receiving subretinal AAV-delivered CRISPR editing for <i>CEP290</i>-associated Leber congenital amaurosis type 10. Preclinical and early clinical studies have further investigated CRISPR-engineered T cells designed to improve antitumor activity, persistence, or resistance to inhibitory signaling. Despite these advances, key translational hurdles include the risk of off-target mutations and large-scale chromosomal rearrangements. Furthermore, immune responses against bacterial Cas9 nucleases and viral delivery vectors may limit the long-term efficacy of CRISPR-based therapies, while technical barriers surrounding delivery to extrahepatic tissues, such as skeletal muscle and the central nervous system, continue to hinder broader clinical success. Ethical concerns regarding germline modifications and the high cost of individualized therapies present additional translational challenges. Consequently, emerging DSB-independent technologies, such as base editing and prime editing, may reduce selected DSB-associated liabilities, but each introduces distinct editing, delivery, and genotoxicity risks that require product-specific evaluation.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1868779"},"PeriodicalIF":5.9,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13601907/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149041581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rewriting the genome: harnessing R2 retrotransposons for precise DNA insertion.","authors":"Liangzheng Fu, Yachao Wu, Xiaohua Jin, Xu Ma","doi":"10.3389/fgeed.2026.1789016","DOIUrl":"10.3389/fgeed.2026.1789016","url":null,"abstract":"<p><p>CRISPR-based genome editors are fundamentally limited by their requirement for double-strand DNA breaks (DSBs), restricted transgene cargo capacity, and reliance on error-prone endogenous DNA repair mechanisms. Non-long terminal repeat (non-LTR) retrotransposons-especially the site-specific R2 element-offer a mechanistically distinct and potentially safer choice for programmable genomic integration. These elements employ target-primed reverse transcription (TPRT)-an RNA-templated integration mechanism that circumvents DSB formation and supports amplification of self-copy. This review delineates the molecular mechanism of R2 retrotransposons, emphasizing their highly specific integration into the 28 S ribosomal DNA locus-a recognized genomic safe harbor. We describe the functional domains of the R2 protein, including the reverse transcriptase, restriction-like endonuclease, and nucleic acid-binding motifs, and explain how they coordinate to achieve precise DNA cleavage and cDNA synthesis. Recent cryo-electron microscopy (cryo-EM) structures have revealed discrete RNA-protein complex that orchestrate the stepwise progression of TPRT. Informed by these mechanistic insights, researchers have engineered programmable platforms-including PRINT and STITCHR-that enable RNA-directed transgene integration in mammalian systems. These platforms establish R2 as a viable all-RNA programmable system for targeted genomic integration. Future directions include reprogramming the DNA-binding specificity of R2 through protein engineering to target loci, optimizing integration fidelity and efficiency, and mining diverse R2-like elements from metagenomic data. With continued optimization and rigorous safety validation, R2-derived platforms could supplant current nuclease-dependent editors in applications requiring high-fidelity, large-cargo integration.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1789016"},"PeriodicalIF":5.9,"publicationDate":"2026-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13586686/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149009377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heat stress impact on rice reproductive processes: challenges and new approaches.","authors":"Miguel Moreira, Ana Rita Queirós, Ana Ventura, Diana Moreira, Sílvia Coimbra, Ana Marta Pereira","doi":"10.3389/fgeed.2026.1808390","DOIUrl":"10.3389/fgeed.2026.1808390","url":null,"abstract":"<p><p>Heat stress represents one of the most severe abiotic constraints to rice (<i>Oryza sativa</i> L.) productivity and is expected to intensify under ongoing climate change, particularly affecting the reproductive phase and leading to substantial yield and grain quality losses. This review synthesizes current knowledge on the impacts of heat stress on rice reproduction, with a focus on both male and female reproductive structures and their interactions. Evidence from anatomical, physiological, transcriptomic, and metabolomic studies to describe how elevated temperatures disrupt key reproductive processes, including microsporogenesis, anther dehiscence, pollen viability, pollen-pistil interactions, fertilisation, and embryo sac development were integrated in this review. It further discusses the genotype-dependent differences in reproductive thermotolerance; and key genes, metabolites, and pathways associated with heat stress perception, signalling, and tolerance are highlighted. Finally, it is briefly discussed how recent advances in breeding strategies, functional genomics and genome-editing technologies, particularly CRISPR-based approaches, are providing new opportunities to enhance reproductive resilience to heat stress and how it is essential to close the existing molecular knowledge gaps in the development of heat-tolerant rice varieties capable of sustaining productivity in a warming climate.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1808390"},"PeriodicalIF":5.9,"publicationDate":"2026-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13548124/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148908343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating high-fidelity CRISPR-Cas nucleases in nucleosomal contexts using a quantitative framework.","authors":"Christopher R Handelmann, Erin Skeens, George P Lisi, Michael J Buck","doi":"10.3389/fgeed.2026.1759382","DOIUrl":"10.3389/fgeed.2026.1759382","url":null,"abstract":"<p><p>Chromatin presents a significant obstacle to CRISPR-Cas gene editing, as chromatin restricts nuclease access to DNA. Recent advances have produced a wide range of high-fidelity Cas9 and Cas12a variants with enhanced properties. However, their precision in targeting DNA within different contexts remains poorly understood. This gap limits our ability to predict and optimize Cas performance in the dynamic chromatin landscape. To elucidate how chromatin variability impacts Cas editing accuracy, we utilized GEMiNI-seq to systematically profile wild-type and engineered Cas9 and Cas12a nucleases across a range of nucleosome sequences. All nucleases showed reduced cleavage in nucleosomal DNA relative to naked DNA, with the strongest inhibition at dyad-proximal sites. Cleavage within nucleosomes was highly variable, with wtSpCas9 exhibiting up to 65-fold different activity depending on the nucleosome type. Editors with high catalytic activity (wtSpCas9, HIFIv2, LbCas12a ULTRA) consistently outperformed high-fidelity variants such as evoSpCas9, which displayed excellent specificity on naked DNA but poor performance in nucleosomal contexts. ROC and PRC analyses revealed that nucleosome sequence and orientation shape both sensitivity and specificity, with HIFIv1 emerging as the top-performing nuclease for nucleosomal targets, while evoSpCas9 excelled in exposed contexts. Our findings demonstrate that local nucleosome sequence and structure profoundly influence Cas nuclease accessibility and specificity. Variability in cleavage across nucleosome types underscores the need to consider chromatin context during target selection and nuclease design. These results provide a framework for selecting or engineering Cas editors optimized for therapeutic genome editing within chromatin.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1759382"},"PeriodicalIF":5.9,"publicationDate":"2026-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13522198/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148852037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Beyond genes: CRISPR/Cas9 for chromosomal engineering in polyploids.","authors":"Mahnoor Jamil, Rizwana Mqabool, Sultan Habibullah Khan, Wang-Qi Huang, Xiu-Hua Chen","doi":"10.3389/fgeed.2026.1916031","DOIUrl":"10.3389/fgeed.2026.1916031","url":null,"abstract":"","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1916031"},"PeriodicalIF":5.9,"publicationDate":"2026-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13522106/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148852018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional genomics-guided design of CAR-T and CAR-NK therapies in hematological malignancies: aligning cellular engineering with immune escape and microenvironmental resistance.","authors":"Shiqi Cai, Yang Tan, Qian Yang, Juan Huang","doi":"10.3389/fgeed.2026.1878460","DOIUrl":"10.3389/fgeed.2026.1878460","url":null,"abstract":"<p><strong>Background: </strong>Chimeric antigen receptor T-cell therapy has changed the treatment landscape of relapsed or refractory hematological malignancies, but primary non-response and post-infusion relapse remain frequent clinical problems. In aggressive B-cell lymphomas, acute leukemias, and multiple myeloma, treatment failure is often driven by overlapping mechanisms rather than a single resistance pathway. These include antigen loss or reduced antigen density, impaired immune recognition, defective inflammatory signaling, checkpoint-mediated suppression, metabolic stress, and limited effector-cell persistence within suppressive disease niches.</p><p><strong>Main body: </strong>Genome engineering has become an important tool for both identifying and addressing these resistance mechanisms. CRISPR-based functional screening, single-cell perturbation approaches, and multi-omics profiling allow immune escape and tumor microenvironment-mediated resistance to be defined more functionally, rather than inferred only from correlative datasets. These insights can inform the design of CAR-T and CAR-NK therapies through multi-target or logic-gated receptors, checkpoint or exhaustion-pathway editing, cytokine-supported and armored constructs, metabolic fitness enhancement, and selected multiplex-editing strategies. In parallel, CAR-NK cells, universal allogeneic CAR-T products, and stem-cell-derived platforms may provide additional options in relapse-prone or heavily pretreated patients, particularly when autologous T-cell fitness, manufacturing feasibility, or repeat dosing is a concern.</p><p><strong>Conclusion: </strong>A resistance-guided approach may help align engineered cellular therapy design with the dominant mechanisms of treatment failure in high-risk hematological malignancies. Rather than simply increasing engineering complexity, future CAR-T and CAR-NK development should link each modification to a measurable resistance mechanism, a feasible biomarker, and a clinically testable benefit. Prospective validation, genomic safety assessment, manufacturing consistency, and long-term monitoring will be essential before resistance-matched cellular immunotherapy can be broadly integrated into clinical practice.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1878460"},"PeriodicalIF":5.9,"publicationDate":"2026-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13519549/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148842288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editorial: Genome editing in poultry and livestock.","authors":"Jiannan Zhang, Lijun Shang","doi":"10.3389/fgeed.2026.1946439","DOIUrl":"https://doi.org/10.3389/fgeed.2026.1946439","url":null,"abstract":"","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1946439"},"PeriodicalIF":5.9,"publicationDate":"2026-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13490097/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148801797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Navigating the human gene editing scene and proposing a new governance framework in Africa: from knowledge to policy to action.","authors":"Abdallah Borham, Fadya M Elgarhy, Aalaa Moghith, Mohamed Tarek Elswefy, Rawan Ibrahim Mansour, May Bakr","doi":"10.3389/fgeed.2026.1866004","DOIUrl":"10.3389/fgeed.2026.1866004","url":null,"abstract":"<p><p>Gene editing holds both hope and despair, necessitating vigilant, ethicolegal oversight to harness its advancement and avoid unintended effects like off-target effects and mosaicism. Despite efforts, governance fragmentation persists regionally and globally. Africa needs a phased, harmonized, continental governance framework for gene editing that combines regional oversight, national implementation, accountability, data governance, genomic sovereignty, and public participation. Policy harmonization is close to accomplishment, but obstacles persist. This paper proposes a practical continent-wide model anchored on African conformity, national legislative capacities, communal engagement, and data sharing. It presents an operational plan encompassing grassroots, nation-state, and regional regulatory bodies, centralized on a new pan-African agency for optimal oversight and governance. The model incorporates bioethical principlism and Ubuntu philosophy, forming the foundations of accountable and safe gene editing research and practice. Trespassing these foundations is a severe, inter-generational, and irresponsible act. This policy review critically analyzes the literature and evidence around gene editing, examining uncertainties, dilemmas, biases, and gaps in concepts. It presents an interpretation of the analysis in a narrative discourse with policy recommendations and governance reforms. The authors relied on empirical evidence and normative arguments. Gene editing is a multifaceted issue, yet a recourse for many patients with devastating diseases. Global progress has been hindered by procedural and operational defects and the controversy of enhancement versus therapy. Declarations, guidelines, and conventions should be prioritized to achieve universal benefits. A diligent consideration of collective risks is crucial to designing effective and inclusive policies. A multi-level approach is imperative for maintaining momentum and pace in technology development and localization.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"8 ","pages":"1866004"},"PeriodicalIF":5.9,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13464102/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148726801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}