Single-molecule live-cell RNA imaging with CRISPR–Csm DOI Creative Commons
Chenglong Xia, David Colognori, X. Jiang

и другие.

Nature Biotechnology, Год журнала: 2025, Номер unknown

Опубликована: Фев. 18, 2025

Abstract Understanding the diverse dynamic behaviors of individual RNA molecules in single cells requires visualizing them at high resolution real time. However, single-molecule live-cell imaging unmodified endogenous has not yet been achieved a generalizable manner. Here, we present fluorescence situ hybridization (smLiveFISH), robust approach that combines programmable RNA-guided, RNA-targeting CRISPR–Csm complex with multiplexed guide RNAs for direct and efficient visualization range cell types, including primary cells. Using smLiveFISH, track native NOTCH2 MAP1B transcripts living identify two distinct localization mechanisms cotranslational translocation mRNA endoplasmic reticulum directional transport toward periphery. This method potential to unlock principles governing spatiotemporal organization health disease.

Язык: Английский

Drug delivery systems for CRISPR-based genome editors DOI
Victoria J. Madigan, Feng Zhang, James E. Dahlman

и другие.

Nature Reviews Drug Discovery, Год журнала: 2023, Номер 22(11), С. 875 - 894

Опубликована: Сен. 18, 2023

Язык: Английский

Процитировано

84
Uncovering the functional diversity of rare CRISPR-Cas systems with deep terascale clustering DOI
Han Altae-Tran, Soumya Kannan,

Anthony J. Suberski

и другие.

Science, Год журнала: 2023, Номер 382(6673)

Опубликована: Ноя. 23, 2023

Microbial systems underpin many biotechnologies, including CRISPR, but the exponential growth of sequence databases makes it difficult to find previously unidentified systems. In this work, we develop fast locality-sensitive hashing-based clustering (FLSHclust) algorithm, which performs deep on massive datasets in linearithmic time. We incorporated FLSHclust into a CRISPR discovery pipeline and identified 188 unreported CRISPR-linked gene modules, revealing additional biochemical functions coupled adaptive immunity. experimentally characterized three HNH nuclease-containing systems, first type IV system with specified interference mechanism, engineered them for genome editing. also candidate VII system, show acts RNA. This work opens new avenues harnessing broader exploration vast functional diversity microbial proteins.

Язык: Английский

Процитировано

80
CRISPR technologies for genome, epigenome and transcriptome editing DOI
Lukas Villiger,

Julia Joung,

Luke W. Koblan

и другие.

Nature Reviews Molecular Cell Biology, Год журнала: 2024, Номер 25(6), С. 464 - 487

Опубликована: Фев. 2, 2024

Язык: Английский

Процитировано

77
Genetics of human brain development DOI
Yi Zhou, Hongjun Song, Guo‐li Ming

и другие.

Nature Reviews Genetics, Год журнала: 2023, Номер 25(1), С. 26 - 45

Опубликована: Июль 28, 2023

Язык: Английский

Процитировано

74
A host of armor: Prokaryotic immune strategies against mobile genetic elements DOI Creative Commons
David Mayo-Muñoz, Rafael Pinilla‐Redondo, Nils Birkholz

и другие.

Cell Reports, Год журнала: 2023, Номер 42(7), С. 112672 - 112672

Опубликована: Июнь 21, 2023

Prokaryotic adaptation is strongly influenced by the horizontal acquisition of beneficial traits via mobile genetic elements (MGEs), such as viruses/bacteriophages and plasmids. However, MGEs can also impose a fitness cost due to their often parasitic nature differing evolutionary trajectories. In response, prokaryotes have evolved diverse immune mechanisms against MGEs. Recently, our understanding abundance diversity prokaryotic systems has greatly expanded. These defense degrade invading material, inhibit genome replication, or trigger abortive infection, leading population protection. this review, we highlight these strategies, focusing on most recent discoveries. The study defenses not only sheds light microbial evolution but uncovers novel enzymatic activities with promising biotechnological applications.

Язык: Английский

Процитировано

69
RNA‐based medicine: from molecular mechanisms to therapy DOI Creative Commons
Anke Sparmann, Jörg Vogel

The EMBO Journal, Год журнала: 2023, Номер 42(21)

Опубликована: Сен. 20, 2023

Abstract RNA‐based therapeutics have the potential to revolutionize treatment and prevention of human diseases. While early research faced setbacks, it established basis for breakthroughs in drug design that culminated extraordinarily fast development mRNA vaccines combat COVID‐19 pandemic. We now reached a pivotal moment where RNA medicines are poised make broad impact clinic. In this review, we present an overview different strategies generate novel therapeutics, including antisense RNAi‐based mechanisms, mRNA‐based approaches, CRISPR‐Cas‐mediated genome editing. Using three rare genetic diseases as examples, highlight opportunities, but also challenges wide‐ranging applications class drugs.

Язык: Английский

Процитировано

60
Chemically Modified Platforms for Better RNA Therapeutics DOI

Yesi Shi,

Xueyan Zhen,

Yiming Zhang

и другие.

Chemical Reviews, Год журнала: 2024, Номер 124(3), С. 929 - 1033

Опубликована: Янв. 29, 2024

RNA-based therapies have catalyzed a revolutionary transformation in the biomedical landscape, offering unprecedented potential disease prevention and treatment. However, despite their remarkable achievements, these encounter substantial challenges including low stability, susceptibility to degradation by nucleases, prominent negative charge, thereby hindering further development. Chemically modified platforms emerged as strategic innovation, focusing on precise alterations either RNA moieties or associated delivery vectors. This comprehensive review delves into platforms, underscoring significance augmenting performance translational prospects of therapeutics. It encompasses an in-depth analysis various chemically that been instrumental propelling therapeutics toward clinical utility. Moreover, scrutinizes rationale behind diverse chemical modification techniques aiming at optimizing therapeutic efficacy molecules, facilitating robust management. Recent empirical studies corroborating enhancement through modifications are highlighted. Conclusively, we offer profound insights transformative impact drugs delineates prospective trajectories for future development integration.

Язык: Английский

Процитировано

53
In vivo editing of lung stem cells for durable gene correction in mice DOI
Yehui Sun, Sumanta Chatterjee,

Xizhen Lian

и другие.

Science, Год журнала: 2024, Номер 384(6701), С. 1196 - 1202

Опубликована: Июнь 13, 2024

In vivo genome correction holds promise for generating durable disease cures; yet, effective stem cell editing remains challenging. this work, we demonstrate that optimized lung-targeting lipid nanoparticles (LNPs) enable high levels of in cells, yielding responses. Intravenously administered gene-editing LNPs activatable tdTomato mice achieved >70% lung editing, sustaining expression >80% epithelial cells 660 days. Addressing cystic fibrosis (CF), NG-ABE8e messenger RNA (mRNA)-sgR553X mediated >95% transmembrane conductance regulator (CFTR) DNA correction, restored CFTR function primary patient-derived bronchial equivalent to Trikafta F508del, corrected intestinal organoids and R553X nonsense mutations 50% CF mice. These findings introduce LNP-enabled tissue disease-modifying correction.

Язык: Английский

Процитировано

50
Application of CRISPR-Cas9 genome editing technology in various fields: A review DOI Creative Commons
Arif Nur Muhammad Ansori, Yulanda Antonius,

Raden JK. Susilo

и другие.

Narra J, Год журнала: 2023, Номер 3(2), С. e184 - e184

Опубликована: Авг. 27, 2023

CRISPR-Cas9 has emerged as a revolutionary tool that enables precise and efficient modifications of the genetic material. This review provides comprehensive overview technology its applications in genome editing. We begin by describing fundamental principles technology, explaining how system utilizes single guide RNA (sgRNA) to direct Cas9 nuclease specific DNA sequences genome, resulting targeted double-stranded breaks. In this review, we provide in-depth explorations agriculture, medicine, environmental sciences, fisheries, nanotechnology, bioinformatics, biotechnology. also highlight potential, ongoing research, ethical considerations controversies surrounding use. might contribute understanding implications various fields, paving way for future developments responsible transformative technology.

Язык: Английский

Процитировано

47
Collateral activity of the CRISPR/RfxCas13d system in human cells DOI Creative Commons
Peiguo Shi, Michael R. Murphy, Alexis O. Aparicio

и другие.

Communications Biology, Год журнала: 2023, Номер 6(1)

Опубликована: Март 28, 2023

Abstract CRISPR/Cas13 systems are increasingly used for programmable targeting of RNAs. While Cas13 nucleases capable degrading both target RNAs and bystander in vitro bacteria, initial studies fail to detect collateral degradation non-target eukaryotic cells. Here we show that RfxCas13d, also known as CasRx, a widely system, can cause transcriptome destruction when abundant reporter RNA endogenous RNAs, resulting proliferation defect these results call caution using RfxCas13d targeted knockdown, demonstrated the activity be harnessed selective depletion specific cell population defined by marker an setting.

Язык: Английский

Процитировано

46