Proceedings of the National Academy of Sciences, Год журнала: 2024, Номер 121(45)
Опубликована: Окт. 30, 2024
CRISPR-Cas13 nucleases are programmable RNA-targeting effectors that can silence gene expression in a transient manner. Recent iterations of Cas13 compact for adeno-associated virus (AAV) delivery to achieve strong and persistent various organs safe Here, we report significant transcriptomic signatures Cas13bt3 retinal cells show all-in-one AAV therapy with effectively
Язык: Английский
Nature Reviews Molecular Cell Biology, Год журнала: 2024, Номер 25(6), С. 464 - 487
Опубликована: Фев. 2, 2024
Язык: Английский
Процитировано
68
Pharmaceutics, Год журнала: 2023, Номер 15(2), С. 685 - 685
Опубликована: Фев. 17, 2023
Retinitis pigmentosa (RP) is a heterogeneous group of hereditary diseases characterized by progressive degeneration retinal photoreceptors leading to visual decline. It the most common type inherited dystrophy and has high burden on both patients society. This condition causes gradual loss vision, with its typical manifestations including nyctalopia, concentric field loss, ultimately bilateral central vision loss. one disability blindness in people under 60 years old affects over 1.5 million worldwide. There currently no curative treatment for RP, only small confirmed RPE65 mutations are eligible receive gene therapy market: voretigene neparvovec. The current therapeutic armamentarium limited retinoids, vitamin A supplements, protection from sunlight, aids, medical surgical interventions treat ophthalmic comorbidities, which aim slow down progression disease. Considering such landscape, there an urgent need developing new individualized modalities targeting degeneration. Although heterogeneity involved RP makes target development difficult, recent fundamental studies showed promising progress elucidation photoreceptor mechanism. discovery novel molecule therapeutics that can selectively specific receptors or pathways will serve as solid foundation advanced drug development. article review focusing preclinical stage research molecular targets, starting point We alterations mainly those regarding endoplasmic reticulum (ER) stress apoptotic pathways, maintenance redox balance, genomic stability. then discuss approaches development, cell therapy, well literature identifying potential targets RP.
Язык: Английский
Процитировано
46
Nature Communications, Год журнала: 2025, Номер 16(1)
Опубликована: Янв. 8, 2025
Abstract An abnormal expansion of a GGGGCC (G 4 C 2 ) hexanucleotide repeat in the C9ORF72 gene is most common genetic cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two debilitating neurodegenerative disorders driven part by gain-of-function mechanisms involving transcribed forms expansion. By utilizing Cas13 variant with reduced collateral effects, we develop here high-fidelity RNA-targeting CRISPR-based system for C9ORF72-linked ALS/FTD. When delivered to brain transgenic rodent model, this Cas13-based platform curbed expression G repeat-containing RNA without affecting normal levels, which turn decreased formation foci, production dipeptide protein, reversed transcriptional deficits. This possessed improved transcriptome-wide specificity compared its native form mediated targeting motor neuron-like cells derived from patient ALS. These results lay foundation implementation CRISPR technologies
Язык: Английский
Процитировано
3
Nature Biotechnology, Год журнала: 2025, Номер unknown
Опубликована: Фев. 11, 2025
Язык: Английский
Процитировано
2
Nature Communications, Год журнала: 2024, Номер 15(1)
Опубликована: Янв. 27, 2024
Abstract Transposon-associated ribonucleoprotein TnpB is known to be the ancestry endonuclease of diverse Cas12 effector proteins from type-V CRISPR system. Given its small size (408 aa), it interest examine whether engineered could used for efficient mammalian genome editing. Here, we showed that gene editing activity native Deinococcus radiodurans (ISDra2 TnpB) in mouse embryos was already higher than previously identified small-sized Cas12f1. Further stepwise engineering noncoding RNA (ωRNA or reRNA) component significantly elevated nuclease TnpB. Notably, an optimized TnpB-ωRNA system efficiently delivered vivo with single adeno-associated virus (AAV) and corrected disease phenotype a tyrosinaemia model. Thus, miniature represents new addition current toolbox, unique feature smallest facilitate AAV delivery cells tissues.
Язык: Английский
Процитировано
12
Molecular Cancer, Год журнала: 2024, Номер 23(1)
Опубликована: Май 15, 2024
Long noncoding RNAs (lncRNAs) have surpassed the number of protein-coding genes, yet majority no known function. We previously discovered 844 lncRNAs that were genetically linked to breast cancer through genome-wide association studies (GWAS). Here, we show a subset these alter risk by modulating cell proliferation, and provide evidence reduced expression on one lncRNA increases aberrant DNA replication repair.
Язык: Английский
Процитировано
10
Nature Communications, Год журнала: 2024, Номер 15(1)
Опубликована: Март 11, 2024
Abstract Current gene silencing tools based on RNA interference (RNAi) or, more recently, clustered regularly interspaced short palindromic repeats (CRISPR)‒Cas13 systems have critical drawbacks, such as off-target effects or collateral mRNA cleavage (CRISPR‒Cas13). Thus, a specific method of knockdown is needed. Here, we develop CRISPRδ, an approach for translational silencing, harnessing catalytically inactive Cas13 proteins (dCas13). Owing to its tight association with mRNA, dCas13 serves physical roadblock scanning ribosomes during translation initiation and does not affect stability. Guide RNAs covering the start codon lead highest efficacy regardless mechanism: cap-dependent, internal ribosome entry site (IRES)-dependent, repeat-associated non-AUG (RAN) translation. Strikingly, genome-wide profiling reveals ultrahigh specificity CRISPRδ. Moreover, fusion repressor further improves performance. Our provides framework repression-based in eukaryotes.
Язык: Английский
Процитировано
9
Cell Communication and Signaling, Год журнала: 2024, Номер 22(1)
Опубликована: Янв. 10, 2024
Abstract Background Claudin-18.2 (CLDN18.2) has emerged as an alluring therapeutic target against gastrointestinal tumors in recent years. However, a thorough understanding of its regulatory mechanism gastric cancer remains elusive. Methods We presented comprehensive study comprising 185 patients, which included 112 cases with high CLDN18.2 expression and 73 low determined by immunohistochemistry. After overdressed AGS NUGC4 cell lines, we elucidated the functions connecting cells cancer-associated fibroblasts (CAFs) through vitro adhesion models vivo lung colonization models. The molecular underlying CLDN18.2-mediated interaction between CAFs was identified RNA sequencing protein-proximity labeling techniques vivo. Results In our own cohort, correlation observed levels advanced stage, poor prognosis, heightened infiltration CAFs. pivotal role mediating CAFs, leads to stroma tissue facilitates clustering into embolus, enhancing cancer’s metastatic progression risk embolic death. Mechanistically, it discovered that can activate metastasis-related signaling pathways CLDN18.2-positive cells. Furthermore, using approach, S100 calcium binding protein A4 (S100A4) distinctive marker interacts enhance progression. Conclusions Our findings illuminated promoting embolism, thereby providing insight potential avenues for positive cancers.
Язык: Английский
Процитировано
8
Nature Communications, Год журнала: 2023, Номер 14(1)
Опубликована: Окт. 14, 2023
The TDP-43 proteinopathies, which include amyotrophic lateral sclerosis and frontotemporal dementia, are a devastating group of neurodegenerative disorders that characterized by the mislocalization aggregation TDP-43. Here we demonstrate RNA-targeting CRISPR effector proteins, programmable class gene silencing agents includes Cas13 family enzymes Cas7-11, can be used to mitigate pathology when programmed target ataxin-2, modifier TDP-43-associated toxicity. In addition inhibiting transit stress granules, find in vivo delivery an ataxin-2-targeting system mouse model proteinopathy improved functional deficits, extended survival, reduced severity neuropathological hallmarks. Further, benchmark platforms against ataxin-2 high-fidelity forms possess transcriptome-wide specificity compared Cas7-11 first-generation effector. Our results potential technology for proteinopathies.
Язык: Английский
Процитировано
19
Nature Communications, Год журнала: 2023, Номер 14(1)
Опубликована: Сен. 20, 2023
Abstract The CRISPR-Cas13 ribonucleases have been widely applied for RNA knockdown and transcriptional modulation owing to their high programmability specificity. However, the large size of Cas13 effectors non-specific cleavage upon target activation limit adeno-associated virus based delivery systems therapeutic applications. Herein, we report detailed biochemical structural characterizations a compact (Cas13bt3) suitable delivery. Distinct from many other systems, Cas13bt3 cleaves nonspecific at internal “UC” sites is activated in length-dependent manner. cryo-electron microscope structure fully active state illustrates basis activation. Guided by structure, obtain engineered variants with minimal off-target yet maintained activities. In conclusion, our data illustrate distinct mechanism guide engineering
Язык: Английский
Процитировано
16