Past, present, and future of CRISPR genome editing technologies

Cell, Journal Year: 2024, Volume and Issue: 187(5), P. 1076 - 1100

Published: Feb. 1, 2024

Genome editing has been a transformative force in the life sciences and human medicine, offering unprecedented opportunities to dissect complex biological processes treat underlying causes of many genetic diseases. CRISPR-based technologies, with their remarkable efficiency easy programmability, stand at forefront this revolution. In Review, we discuss current state CRISPR gene technologies both research therapy, highlighting limitations that constrain them technological innovations have developed recent years address them. Additionally, examine summarize landscape applications context health therapeutics. Finally, outline potential future developments could shape coming years.

Language: Английский

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Structural mechanism of bridge RNA-guided recombination

Nature, Journal Year: 2024, Volume and Issue: 630(8018), P. 994 - 1002

Published: June 26, 2024

Insertion sequence (IS) elements are the simplest autonomous transposable found in prokaryotic genomes

Language: Английский

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Rapid DNA unwinding accelerates genome editing by engineered CRISPR-Cas9

Cell, Journal Year: 2024, Volume and Issue: 187(13), P. 3249 - 3261.e14

Published: May 22, 2024

Thermostable clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas9) enzymes could improve genome-editing efficiency delivery due to extended protein lifetimes. However, initial experimentation demonstrated Geobacillus stearothermophilus Cas9 (GeoCas9) be virtually inactive when used in cultured human cells. Laboratory-evolved variants of GeoCas9 overcome this natural limitation by acquiring mutations the wedge (WED) domain that produce >100-fold-higher levels. Cryoelectron microscopy (cryo-EM) structures wild-type improved (iGeoCas9) reveal contacts between WED iGeoCas9 DNA substrates. Biochemical analysis shows accelerates unwinding capture substrates under magnesium-restricted conditions typical mammalian but not bacterial These findings enabled rational engineering other orthologs enhance levels, pointing a general strategy for editing enzyme improvement. Together, these results uncover new role demonstrate how accelerated target dramatically improves Cas9-induced activity.

Language: Английский

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Transposon-encoded nucleases use guide RNAs to promote their selfish spread

Nature, Journal Year: 2023, Volume and Issue: 622(7984), P. 863 - 871

Published: Sept. 27, 2023

Language: Английский

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CRISPR/Cas9 Landscape: Current State and Future Perspectives

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(22), P. 16077 - 16077

Published: Nov. 8, 2023

CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 is a unique genome editing tool that can be easily used in wide range of applications, including functional genomics, transcriptomics, epigenetics, biotechnology, plant engineering, livestock breeding, gene therapy, diagnostics, and so on. This review focused on the current CRISPR/Cas9 landscape, e.g., Cas9 variants with improved properties, Cas9-derived fusion proteins, delivery methods, pre-existing immunity against anti-CRISPR their possible roles function improvement. Moreover, this presents detailed outline CRISPR/Cas9-based diagnostics therapeutic approaches. Finally, addresses future expansion editors’ toolbox orthologs other CRISPR/Cas proteins.

Language: Английский

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Programmable RNA-guided DNA endonucleases are widespread in eukaryotes and their viruses

Science Advances, Journal Year: 2023, Volume and Issue: 9(39)

Published: Sept. 27, 2023

Programmable RNA-guided DNA nucleases perform numerous roles in prokaryotes, but the extent of their spread outside prokaryotes is unclear. Fanzors, eukaryotic homolog prokaryotic TnpB proteins, have been detected genomes eukaryotes and large viruses, activity functions remain unknown. Here, we characterize Fanzors as RNA-programmable endonucleases, using biochemical cellular evidence. We found diverse that frequently associate with various transposases. Reconstruction evolution revealed multiple radiations RuvC-containing homologs eukaryotes. Fanzor genes captured introns proteins acquired nuclear localization signals, indicating extensive, long-term adaptation to functioning cells. contain a rearranged catalytic site RuvC domain, similar distinct subset TnpBs, lack collateral cleavage activity. demonstrate can be harnessed for genome editing human cells, highlighting potential these widespread biotechnology applications.

Language: Английский

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GMOs or non-GMOs? The CRISPR Conundrum

Frontiers in Plant Science, Journal Year: 2023, Volume and Issue: 14

Published: Oct. 9, 2023

CRISPR-Cas9, the “genetic scissors”, is being presaged as a revolutionary technology, having tremendous potential to create designer crops by introducing precise and targeted modifications in genome achieve global food security face of climate change increasing population. Traditional genetic engineering relies on random unpredictable insertion isolated genes or foreign DNA elements into plant genome. However, CRISPR-Cas based gene editing does not necessarily involve inserting element from different species but new traits precisely altering existing genes. CRISPR edited are touching markets, however, world community divided over whether these should be considered genetically modified (GM) non-GM. Classification crops, especially transgene free traditional GM will significantly affect their future public acceptance some regions. Therefore, depending upon regulation non-GMs, perception. Here we briefly discuss how crops. In addition, reagents delivery tools produce transgene-free Moreover, also summarize regulatory classification countries regulating We that controversy CRISPR-edited plants non-GM continue until universal, transparent, scalable framework for introduced worldwide, with increased awareness involving all stakeholders.

Language: Английский

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CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane

Frontiers in Plant Science, Journal Year: 2024, Volume and Issue: 14

Published: Jan. 8, 2024

Gene editing technologies have opened up the possibility of manipulating genome any organism in a predicted way. CRISPR technology is most used tool and, agriculture, it has allowed expansion possibilities plant biotechnology, such as gene knockout or knock-in, transcriptional regulation, epigenetic modification, base editing, RNA prime and nucleic acid probing detection. This mostly depends on

Language: Английский

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Emerging Gene Therapeutics for Epidermolysis Bullosa under Development

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(4), P. 2243 - 2243

Published: Feb. 13, 2024

The monogenetic disease epidermolysis bullosa (EB) is characterised by the formation of extended blisters and lesions on patient's skin upon minimal mechanical stress. Causal for this severe condition are genetic mutations in genes, leading to functional impairment, reduction, or absence encoded protein within skin's basement membrane zone connecting epidermis underlying dermis. major burden affected families justifies development long-lasting curative therapies operating at genomic level. landscape causal EB steadily expanding due recent breakthroughs gene therapy field, providing promising outcomes patients suffering from disease. Currently, two therapeutic approaches show promise EB. clinically more advanced replacement strategy was successfully applied forms, a ground-breaking vivo product named beremagene geperpavec (B-VEC) recently approved US Food Drug Administration (FDA). In addition, continuous innovations both designer nucleases editing technologies enable efficient potentially safe repair permanent manner, inspiring researchers field define reach new milestones

Language: Английский

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Beyond the promise: evaluating and mitigating off-target effects in CRISPR gene editing for safer therapeutics

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 11

Published: Jan. 18, 2024

Over the last decade, CRISPR has revolutionized drug development due to its potential cure genetic diseases that currently do not have any treatment. was adapted from bacteria for gene editing in human cells 2012 and, remarkably, only 11 years later seen it’s very first approval as a medicine treatment of sickle cell disease and transfusion-dependent beta-thalassemia. However, application systems is associated with unintended off-target on-target alterations (including small indels, structural variations such translocations, inversions large deletions), which are source risk patients vital concern safe therapies. In recent years, wide range methods been developed detect unwanted effects CRISPR-Cas nuclease activity. this review, we summarize different assessment, discuss their strengths limitations, highlight strategies improve safety systems. Finally, relevance pre-clinical assessment therapeutics within current regulatory context.

Language: Английский

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