Navigating the CRISPR/Cas Landscape for Enhanced Diagnosis and Treatment of Wilson’s Disease DOI Creative Commons
Woong Choi, Seongkwang Cha, Kyoungmi Kim

и другие.

Cells, Год журнала: 2024, Номер 13(14), С. 1214 - 1214

Опубликована: Июль 18, 2024

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system continues to evolve, thereby enabling more precise detection and repair of mutagenesis. development CRISPR/Cas-based diagnosis holds promise for high-throughput, cost-effective, portable nucleic acid screening genetic disease diagnosis. In addition, advancements in transportation strategies such as adeno-associated virus (AAV), lentiviral vectors, nanoparticles, virus-like vectors (VLPs) offer synergistic insights gene therapeutics vivo. Wilson's (WD), a copper metabolism disorder, is primarily caused by mutations the ATPase transporting beta (

Язык: Английский

Systematic optimization of prime editing for the efficient functional correction of CFTR F508del in human airway epithelial cells DOI Creative Commons

Alexander A. Sousa,

Colin Hemez, Lei Lei

и другие.

Nature Biomedical Engineering, Год журнала: 2024, Номер unknown

Опубликована: Июль 10, 2024

Abstract Prime editing (PE) enables precise and versatile genome without requiring double-stranded DNA breaks. Here we describe the systematic optimization of PE systems to efficiently correct human cystic fibrosis (CF) transmembrane conductance regulator ( CFTR ) F508del, a three-nucleotide deletion that is predominant cause CF. By combining six efficiency optimizations for PE—engineered guide RNAs, PEmax architecture, transient expression dominant-negative mismatch repair protein, strategic silent edits, PE6 variants proximal ‘dead’ single-guide RNAs—we increased correction efficiencies F508del from less than 0.5% in HEK293T cells 58% immortalized bronchial epithelial (a 140-fold improvement) 25% patient-derived airway cells. The also resulted minimal off-target editing, edit-to-indel ratios 3.5-fold greater those achieved by nuclease-mediated homology-directed repair, functional restoration ion channels over 50% wild-type levels (similar via combination treatment with elexacaftor, tezacaftor ivacaftor) primary Our findings support feasibility durable one-time

Язык: Английский

Процитировано

23
Leveraging CRISPR gene editing technology to optimize the efficacy, safety and accessibility of CAR T-cell therapy DOI Creative Commons
Tao Lei, Yazhuo Wang, Yuchen Zhang

и другие.

Leukemia, Год журнала: 2024, Номер unknown

Опубликована: Окт. 25, 2024

Язык: Английский

Процитировано

18
Recent Therapeutic Gene Editing Applications to Genetic Disorders DOI Creative Commons
Éric Deneault

Current Issues in Molecular Biology, Год журнала: 2024, Номер 46(5), С. 4147 - 4185

Опубликована: Апрель 30, 2024

Recent years have witnessed unprecedented progress in therapeutic gene editing, revolutionizing the approach to treating genetic disorders. In this comprehensive review, we discuss progression of milestones leading emergence clustered regularly interspaced short palindromic repeats (CRISPR)-based technology as a powerful tool for precise and targeted modifications human genome. CRISPR-Cas9 nuclease, base prime editing taken center stage, demonstrating remarkable precision efficacy ex vivo genomic modifications. Enhanced delivery systems, including viral vectors nanoparticles, further improved efficiency safety advancing their clinical translatability. The exploration CRISPR-Cas systems beyond commonly used Cas9, such development Cas12 Cas13 variants, has expanded repertoire tools, enabling more intricate interventions. Outstandingly, represents significant leap forward, given its unparalleled versatility minimization off-target effects. These innovations paved way multitude previously incurable disorders, ranging from monogenic diseases complex polygenic conditions. This review highlights latest innovative studies field, emphasizing breakthrough technologies preclinical trials, applications realm medicine. However, challenges effects ethical considerations remain, necessitating continued research refine profiles frameworks.

Язык: Английский

Процитировано

13
Open-ended molecular recording of sequential cellular events into DNA DOI
Theresa B. Loveless, Courtney Carlson,

Catalina A. Dentzel Helmy

и другие.

Nature Chemical Biology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 14, 2024

Язык: Английский

Процитировано

10
Randomizing the human genome by engineering recombination between repeat elements DOI
Jonas Koeppel, Raphaël Ferreira,

Thomas Vanderstichele

и другие.

Science, Год журнала: 2025, Номер 387(6733)

Опубликована: Янв. 30, 2025

We lack tools to edit DNA sequences at scales necessary study 99% of the human genome that is noncoding. To address this gap, we applied CRISPR prime editing insert recombination handles into repetitive sequences, up 1697 per cell line, which enables generating large-scale deletions, inversions, translocations, and circular DNA. Recombinase induction produced more than 100 stochastic megabase-sized rearrangements in each cell. tracked these over time measure selection pressures, finding a preference for shorter variants avoided essential genes. characterized 29 clones with multiple rearrangements, an impact deletions on expression genes variant but not nearby This genome-scrambling strategy large sequence relocations, insertion regulatory elements explore dispensability organization.

Язык: Английский

Процитировано

2
tRNA-derived small RNAs: their role in the mechanisms, biomarkers, and therapeutic strategies of colorectal cancer DOI Creative Commons
Bo Zhang, Yi Pan, Zhe Li

и другие.

Journal of Translational Medicine, Год журнала: 2025, Номер 23(1)

Опубликована: Янв. 13, 2025

Colorectal cancer (CRC) is the third most prevalent malignancy and second leading cause of cancer-related mortality worldwide, with an increasing shift towards younger age onset. In recent years, there has been recognition significance tRNA-derived small RNAs (tsRNAs), encompassing fragments (tRFs) tRNA halves (tiRNAs). Their involvement in regulating translation, gene expression, reverse transcription, epigenetics gradually come to light. Emerging research revealed dysregulation tsRNAs CRC, implicating their role CRC initiation progression, highlighting potential early diagnosis, prognosis, therapeutic strategies. Although clinical application still its stages, findings highlight a close relationship between biogenesis function tsRNAs, chemical modifications, tumor immune microenvironment (TIME). Additionally, similar other RNAs, can be effectively delivered via nanoparticles (NPs). Consequently, future should focus on elucidating concerning base TIME regulation, immunotherapy, NPs delivery systems facilitate translation.

Язык: Английский

Процитировано

1
EXPERT expands prime editing efficiency and range of large fragment edits DOI Creative Commons
Youcai Xiong, Yuhong Su,

Ruigao He

и другие.

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Фев. 12, 2025

Abstract Prime editing systems (PEs) hold great promise in modern biotechnology. However, their range is limited as PEs can only modify the downstream sequences of pegRNA nick. Here, we report development ex tended p rime e dito r sys t em (EXPERT) to overcome this limitation by using an extended (ext-pegRNA) with modified 3’ extension, and additional sgRNA (ups-sgRNA) targeting upstream region ext-pegRNA. We demonstrate that EXPERT efficiently perform on both sides ext-pegRNA nick, a task unattainable canonical PEs. exhibits prominent capacity replacing up 88 base pairs inserting 100 within Compared such PE2, utilization strategy significantly enhances efficiency for large fragment edits average improvement 3.12-fold, 122.1 times higher. Safety wise, use ups-sgRNA does not increase rates undesirable insertions deletions (indels), two nicks are same strand. Moreover, do observe increased off-target genome-wide. Our work introduces PE tool significant potential life sciences.

Язык: Английский

Процитировано

1
Navigating gene editing in porcine embryos: Methods, challenges, and future perspectives DOI Creative Commons
Julieta Gabriela Hamze, Josep M. Cambra, Sergio Navarro-Serna

и другие.

Genomics, Год журнала: 2025, Номер 117(2), С. 111014 - 111014

Опубликована: Фев. 13, 2025

Gene editing technologies, particularly CRISPR/Cas9, have emerged as transformative tools in genetic modification, significantly advancing the use of porcine embryos biomedical and agricultural research. This review comprehensively examines various methodologies for gene delivery methods, such somatic cell nuclear transfer (SCNT), microinjection, electroporation, lipofection. review, focuses on advantages or limitations using different biological sources (in vivo- vs. vitro oocytes/embryos). Male germ manipulation sperm-mediated (SMGT) testis-mediated (TMGT) represent innovative approaches producing genetically modified animals. Although these technologies revolutionized engineering field, all strategies face challenges, including high rates off-target events mosaicism. emphasizes need to refine with a focus reducing mosaicism improving accuracy. Further advancements are essential unlocking full potential both applications innovations.

Язык: Английский

Процитировано

1
Prime Editing: Mechanistic Insights and DNA Repair Modulation DOI Creative Commons

Astrid Mentani,

Marcello Maresca, Anna Shiriaeva

и другие.

Cells, Год журнала: 2025, Номер 14(4), С. 277 - 277

Опубликована: Фев. 13, 2025

Prime editing is a genome technique that allows precise modifications of cellular DNA without relying on donor templates. Recently, several different prime editor proteins have been published in the literature, single- or double-strand breaks. When occurs, undergoes one repair pathways, and these processes can be modulated with use inhibitors. Firstly, this review provides an overview mechanisms their modulation by known In addition, we summarize editors provide comprehensive associated mechanisms. Finally, discuss delivery safety aspects editing.

Язык: Английский

Процитировано

1
Emerging applications of gene editing technologies for the development of climate-resilient crops DOI Creative Commons
R. L. Chavhan,

Siddhant Gahininath Jaybhaye,

V. R. Hinge

и другие.

Frontiers in Genome Editing, Год журнала: 2025, Номер 7

Опубликована: Март 10, 2025

Climate change threatens global crop yield and food security due to rising temperatures, erratic rainfall, increased abiotic stresses like drought, heat, salinity. Gene editing technologies, including CRISPR/Cas9, base editors, prime offer precise tools for enhancing resilience. This review explores the mechanisms of these technologies their applications in developing climate-resilient crops address future challenges. While CRISPR/enables targeted modifications plant DNA, editors allow direct conversion without inducing double-stranded breaks, enable insertions, deletions, substitutions. By understanding manipulating key regulator genes involved stress responses, such as DREB, HSP, SOS, ERECTA, HsfA1, NHX; tolerance can be enhanced against salt stress. improve traits related root development, water use efficiency, response pathways, heat shock response, photosynthesis, membrane stability, ion homeostasis, osmotic adjustment, oxidative response. Advancements gene integration with genomics, phenomics, artificial intelligence (AI)/machine learning (ML) hold great promise. However, challenges off-target effects, delivery methods, regulatory barriers must addressed. highlights potential develop crops, contributing sustainable agriculture.

Язык: Английский

Процитировано

1