Orthogonal and multiplexable genetic perturbations with an engineered prime editor and a diverse RNA array DOI Creative Commons
Qichen Yuan, Hongzhi Zeng, T. Daniel

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Дек. 30, 2024

Programmable and modular systems capable of orthogonal genomic transcriptomic perturbations are crucial for biological research treating human genetic diseases. Here, we present the minimal versatile perturbation technology (mvGPT), a flexible toolkit designed simultaneous gene editing, activation, repression in cells. The mvGPT combines an engineered compact prime editor (PE), fusion activator MS2-p65-HSF1 (MPH), drive-and-process multiplex array that produces RNAs tailored to different types perturbation. can precisely edit genome via PE coupled with editing guide RNA nicking RNA, activate endogenous expression using truncated single containing MPH-recruiting MS2 aptamers, silence interference short-hairpin RNA. We showcase versatility by simultaneously correcting c.3207C>A mutation ATP7B linked Wilson's disease, upregulating PDX1 potentially treat Type I diabetes, suppressing TTR manage transthyretin amyloidosis. In addition plasmid delivery, successfully utilize various methods deliver payload, demonstrating its potential future vivo applications.

Язык: Английский

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
CRISPR/Cas9 Genome Editing in the Diamondback Moth: Current Progress, Challenges, and Prospects DOI Open Access
Muhammad Asad,

Yanpeng Chang,

Jianying Liao

и другие.

International Journal of Molecular Sciences, Год журнала: 2025, Номер 26(4), С. 1515 - 1515

Опубликована: Фев. 11, 2025

The development of site-specific genome-editing tools like CRISPR (clustered regularly interspaced short palindromic repeat) and its associated protein, Cas9, is revolutionizing genetic engineering with highly efficient mechanism, offering the potential for effective pest management. Recently, CRISPR/Cas9 gene-editing has been extensively utilized in management diamondback moth, Plutella xylostella (L.), a destructive vegetable crops, different purposes, such as gene function analysis control. However, progress related to this tool P. not yet summarized. This review highlights applications uncovering genes critical development, reproduction, insecticide resistance xylostella. Moreover, drive population suppression modifications also discussed. In addition significant made, challenges low germline editing efficiency limited homology-directed repair remain obstacles widespread application. To address these limitations, we have discussed strategies that are anticipated improve CRISPR/Cas9, paving way it becoming pivotal sustainable Therefore, present will help researchers future enhance system use manage moth.

Язык: Английский

Процитировано

0
Make-or-break prime editing for genome engineering in Streptococcus pneumoniae DOI Creative Commons

Monica Rengifo-Gonzalez,

Maria-Vittoria Mazzuoli, Axel B. Janssen

и другие.

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

Опубликована: Апрель 23, 2025

Язык: Английский

Процитировано

0
Make-or-break prime editing for bacterial genome engineering DOI Creative Commons

Monica Rengifo-Gonzalez,

Maria-Vittoria Mazzuoli, Axel B. Janssen

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 28, 2024

Abstract CRISPR-Cas9 has revolutionized genome engineering by allowing precise introductions of DNA double-strand breaks (DSBs). However, in bacteria is still a complex, multi-step process requiring donor template for homology-directed repair DSBs. Prime editing circumvents this need as the provided within prime guide RNA (pegRNA). Here, we developed make-or-break Editing (mbPE) that allows and effective genetic opportunistic human pathogen Streptococcus pneumoniae . In contrast to traditional which nicking Cas9 employed, mbPE harnesses wild type combination with pegRNA destroys seed region or protospacer adjacent motif. Since most poorly perform template-independent end joining, correctly genome-edited clones are selectively enriched during mbPE. We show can be used introduce point mutations, deletions targeted insertions, including protein tags such split luciferase, at selection efficiencies over 93%. enables sequential editing, scalable, generate pools mutants high-throughput manner. The system design guidelines described here will ameliorate future bacterial endeavors.

Язык: Английский

Процитировано

2
State of the art CRISPR-based strategies for cancer diagnostics and treatment DOI Creative Commons
Emma Di Carlo, Carlo Sorrentino

Biomarker Research, Год журнала: 2024, Номер 12(1)

Опубликована: Дек. 18, 2024

Abstract Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology is a groundbreaking and dynamic molecular tool for DNA RNA “surgery”. CRISPR/Cas9 the most widely applied system in oncology research. It major advancement genome manipulation due to its precision, efficiency, scalability versatility compared previous gene editing methods. has shown great potential not only targeting of oncogenes or genes coding immune checkpoint molecules, engineering T cells, but also epigenomic disturbances, which contribute cancer development progression. proven useful detecting genetic mutations, enabling large-scale screening involved tumor onset, progression drug resistance, speeding up highly targeted therapies tailored immunological profiles patient’s tumor. Furthermore, recently discovered Cas12 Cas13 systems have expanded Cas9-based applications, providing new opportunities diagnosis treatment cancer. In addition traditional cis-cleavage, they exhibit trans-cleavage activity, enables their use as sensitive specific diagnostic tools. Diagnostic platforms like DETECTR, employs enzyme, that cuts single-stranded reporters, SHERLOCK, uses Cas12, Cas13, specifically target cleave RNA, can be exploited speed advance oncological diagnostics. Overall, CRISPR platform improve diagnostics functionality safety engineered cellular medicines. Here, we will emphasize potentially transformative impact field treatments, prognostic approaches, highlight challenges raised by using newly introduced CRISPR-based therapy.

Язык: Английский

Процитировано

2
Efficient Genome Editing with Chimeric Oligonucleotide-Directed Editing DOI
Long Thành Nguyễn, Noah R. Rakestraw,

Brianna L.M. Pizzano

и другие.

Опубликована: Июль 10, 2024

Abstract Prime editing has emerged as a precise and powerful genome tool, offering favorable gene profile compared to other Cas9-based approaches. Here we report new nCas9-DNA polymerase fusion proteins create chimeric oligonucleotide-directed (CODE) systems for search-and-replace editing. Through successive rounds of engineering, developed CODEMax CODEMax(exo+) editors that achieve efficient modifications in human cells with low unintended edits. contain an engineered Bst DNA derivative known its robust strand displacement ability. Additionally, features 5’ 3’ exonuclease activity promotes effective invasion repair outcomes favoring the incorporation desired edit. We demonstrate CODEs can perform small insertions, deletions, substitutions improved efficiency PEMax at many loci. Overall, complement existing prime expand toolbox manipulations without double-stranded breaks.

Язык: Английский

Процитировано

1
The future of complement therapeutics DOI Creative Commons
Martin Kolev,

Kollu N. Rao,

Michael W. Yeh

и другие.

Exploration of Immunology, Год журнала: 2024, Номер unknown, С. 577 - 615

Опубликована: Окт. 18, 2024

Complement is both evolutionary and scientifically old. It predates the adaptive immunity by some 600 million years was first described in 1905 Jules Bordet Paul Ehrlich. For most of its, existence complement system has been ignored scientists clinicians due to perception it being complicated its relevance for pathogenesis human disease unclear. With recent US Food Drug Administration (FDA) approvals pegcetacoplan paroxysmal nocturnal haemoglobinuria (PNH) geographic atrophy (GA), avacincaptad pegol GA iptacopan danicopan PNH, we are at a crucial juncture complement-targeting therapies. A number companies academic institutions developing next-generation therapies, which resulting an increasingly competitive landscape. If one looks serum cascade, all 3 pathways now have biotechnology or pharmaceutical industry players with 1 multiple clinical-stage inhibitors that expected be FDA approved within next few years. Furthermore, limited clinically validated targets complement-mediated disease, competition this space set further intensify coming In review, will discuss timeline discoveries led development current crop FDA-approved therapeutics. We follow discussion crowded therapy scientific advances emerged two decades underpinning future innovation, including our understanding biology, such as local intracellular complement, emerging targets, combinational approaches non-complement therapeutics unlock new indications technologies gene therapy. also give comprehensive overview landscape how can utilized target dysregulation.

Язык: Английский

Процитировано

1
Bidirectional Prime Editing: Combining Precision with Versatility for Genome Editing DOI
Mahmood S. Choudhery, Taqdees Arif,

Ruhma Mahmood

и другие.

Cellular Reprogramming, Год журнала: 2024, Номер unknown

Опубликована: Дек. 17, 2024

Genome editing techniques have potential to revolutionize the field of life sciences. Several limitations associated with traditional gene been resolved development prime editors that precisely edit DNA without double-strand breaks (DSBs). To further improve efficiency, several modified versions (PE) system introduced. Bi-directional PE (Bi-PE), for example, uses two guide RNAs enabling broad and improved efficiency. It has alter, delete, integrate, replace larger genome sequences multiple bases at same time. This review aims discuss typical methods offer DSB-mediated repair mechanisms, followed by latest advances in technologies non-DSB-mediated repair. The specifically focuses on Bi-PE being an efficient tool human genome. In addition, discusses applications, limitations, future perspectives editing.

Язык: Английский

Процитировано

1
From pollutants to products: Microbial cell factories driving sustainable biomanufacturing and environmental conservation DOI
Maria Gavrilescu

Chemical Engineering Journal, Год журнала: 2024, Номер unknown, С. 157152 - 157152

Опубликована: Окт. 1, 2024

Язык: Английский

Процитировано

0
Orthogonal and multiplexable genetic perturbations with an engineered prime editor and a diverse RNA array DOI Creative Commons
Qichen Yuan, Hongzhi Zeng, T. Daniel

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Дек. 30, 2024

Programmable and modular systems capable of orthogonal genomic transcriptomic perturbations are crucial for biological research treating human genetic diseases. Here, we present the minimal versatile perturbation technology (mvGPT), a flexible toolkit designed simultaneous gene editing, activation, repression in cells. The mvGPT combines an engineered compact prime editor (PE), fusion activator MS2-p65-HSF1 (MPH), drive-and-process multiplex array that produces RNAs tailored to different types perturbation. can precisely edit genome via PE coupled with editing guide RNA nicking RNA, activate endogenous expression using truncated single containing MPH-recruiting MS2 aptamers, silence interference short-hairpin RNA. We showcase versatility by simultaneously correcting c.3207C>A mutation ATP7B linked Wilson's disease, upregulating PDX1 potentially treat Type I diabetes, suppressing TTR manage transthyretin amyloidosis. In addition plasmid delivery, successfully utilize various methods deliver payload, demonstrating its potential future vivo applications.

Язык: Английский

Процитировано

0