The Continuous Improvement of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)–CRISPR-Associated Protein System Has Led to Its Highly Efficient Application in Plants DOI Creative Commons

W. Z. Tan,

Zhiyuan Wang, Liezhao Liu

et al.

Agriculture, Journal Year: 2024, Volume and Issue: 15(1), P. 29 - 29

Published: Dec. 26, 2024

The creation of the CRISPR–Cas system has provided unprecedented opportunities in plant genome research and crop genetic improvement. In recent years, this been continuously improved to meet human needs through expansion modification Cas proteins, diversification targeting locations, optimization CRISPR vectors. review, we systematically describe Class II proteins that have used plants, deactivated Cas9 (dCas9) its role transcriptional regulation, precision editing systems, protein variants, as well methods examples systems various regions with different breadths. addition, outline plans based on constructs can overcome pleiotropy genes or accelerate generation transgene-free plants applications breeding practices. Finally, discuss theory development “CRISPR plus”, integrated application existing more species.

Language: Английский

CRISPR–Cas applications in agriculture and plant research DOI
Aytug Tuncel, Changtian Pan,

Joshua S. Clem

et al.

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 7, 2025

Language: Английский

Citations

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

et al.

Frontiers in Genome Editing, Journal Year: 2025, Volume and Issue: 7

Published: March 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.

Language: Английский

Citations

1
An improved plant prime editor for efficient generation of multiple-nucleotide variations and structural variations in rice DOI Creative Commons
Zhaohui Zhong, Tingting Fan, Yao He

et al.

Plant Communications, Journal Year: 2024, Volume and Issue: 5(9), P. 100976 - 100976

Published: May 14, 2024

Language: Английский

Citations

6
Genome-Wide A → G and C → T Mutations Induced by Functional TadA Variants in Escherichia coli DOI
Hao Wang,

Zhengxin Dong,

Jingyi Shi

et al.

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 9, 2025

The fusion expression of deoxyribonucleic acid (DNA) replication-related proteins with nucleotide deaminase enzymes promotes random mutations in bacterial genomes, thereby increasing genetic diversity among the population. Most previous studies have focused on cytosine deaminase, which produces only C → T mutations, significantly limiting variety mutation types. In this study, we developed a system by combining DnaG (RNA primase) adenine TadA-8e (DnaG-TadA) Escherichia coli, is capable rapidly introducing A G into E. coli genome, resulting 664-fold increase terms rate. Additionally, tested dual-functional TadA variant, TadAD, and then fused it DnaG. This construct introduced both rate increased 370-fold upon coexpression uracil glycosylase inhibitor (DnaG-TadAD-UGI). We applied DnaG-TadA DnaG-TadAD-UGI systems to adaptive laboratory evolution for Cd2+ kanamycin resistance, achieving an 8.0 mM 200 μg/mL tolerance within just 17 days 132 h, respectively. Compared conventional methods, final levels were 320 266%, Our work offers novel strategy mutagenesis potentially other prokaryotic species.

Language: Английский

Citations

0
Harnessing novel cytidine deaminases from the animal kingdom for robust multiplexed base editing in rice DOI Creative Commons
Danyel Fernandes Contiliani, Simon Sretenovic,

Micah Dailey

et al.

Plant Biotechnology Journal, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 14, 2025

CRISPR-Cas-based cytosine base editors (CBEs) are prominent tools that perform site-specific and precise C-to-T conversions catalysed by cytidine deaminases. However, their use is often constrained stringent editing preferences for genomic contexts, off-target effects restricted windows. To expand the repertoire of CBEs, we systematically screened 66 novel deaminases sourced from various organisms, predominantly animal kingdom benchmarked them in rice protoplasts using nCas9-BE3 configuration. After selecting candidates further validation transgenic lines, unveiled a few exhibiting high efficiencies wide CBEs based on these also displayed minimal frequencies indels C-to-R (R = A/G) conversions, suggesting purity editing. Furthermore, highlight highly efficient deaminase OoA3GX2 derived Orca (killer whale) its comparable activity across GC/CC/TC/AC sites, thus broadening targeting scope robust multiplexed Finally, whole-genome sequencing analyses revealed very sgRNA-dependent -independent independent T0 lines. This study expands base-editing toolkit with many mammals, providing better-performing can be leveraged sophisticated genome engineering strategies likely other plant species.

Language: Английский

Citations

0
Examining the adsorption and sensing characteristics of cytosine (CTE) on Y9N9 (Y = Al, B, Ga) nanorings using solvent effects, DFT, AIM and SERS analyses DOI
Jamelah S. Al‐Otaibi, Y. Sheena Mary,

Unnati Jethawa

et al.

Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy, Journal Year: 2025, Volume and Issue: unknown, P. 126148 - 126148

Published: March 1, 2025

Language: Английский

Citations

0
Improving plant C-to-G base editors with a cold-adapted glycosylase and TadA-8e variants DOI
Yingli Jiang, Xiao Zhi,

Zhaopeng Luo

et al.

Trends in biotechnology, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Language: Английский

Citations

0
Versatile plant genome engineering using anti-CRISPR-Cas12a systems DOI
Yao He, Shishi Liu, Long Chen

et al.

Science China Life Sciences, Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 15, 2024

Language: Английский

Citations

2
An efficient CRISPR‐Cas12a‐mediated MicroRNA knockout strategy in plants DOI Creative Commons
Xuelian Zheng, Xu Tang, Yuechao Wu

et al.

Plant Biotechnology Journal, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 14, 2024

Summary In recent years, the CRISPR‐Cas9 nuclease has been used to knock out MicroRNA (miRNA) genes in plants, greatly promoting study of miRNA function. However, due its propensity for generating small insertions and deletions, Cas9 is not well‐suited achieving a complete knockout genes. By contrast, CRISPR‐Cas12a generates larger which could significantly disrupt secondary structure pre‐miRNA prevent production mature miRNAs. Through case OsMIR390 rice, we confirmed that Cas12a more efficient tool than mutants gene. To further demonstrate CRISPR‐Cas12a‐mediated targeted nine OsMIRNA have different spaciotemporal expression previously investigated via genetic approaches. With CRISPR‐Cas12a, up 100% genome editing efficiency was observed at these loci. The resulting deletions suggest robustly generated null alleles Transcriptome profiling mutants, as well phenotypic analysis rice grains revealed function miRNAs controlling gene regulating grain quality seed development. This established an plants.

Language: Английский

Citations

2
In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy DOI Creative Commons
Weiyue Zhang, Xin Huang

Journal of Hematology & Oncology, Journal Year: 2024, Volume and Issue: 17(1)

Published: Nov. 13, 2024

Chimeric antigen receptor (CAR) cell therapy has achieved groundbreaking success in treating hematological malignancies. However, its application to solid tumors remains challenging due complex manufacturing processes, limited vivo persistence, and transient therapeutic effects. In CAR-immune cells induced by gene delivery systems loaded with CAR genes gene-editing tools have shown efficiency for anti-tumor immunotherapy. situ programming of autologous immune avoids the safety concerns allogeneic cells, manufacture could be standardized. Therefore, editing generation might potentially overcome abovementioned limitations current therapy. This review mainly focuses on structures, tools, techniques applied immunotherapy help design develop The recent applications both hematologic malignancies are investigated. To sum up, holds promise offering a practical, cost-effective, efficient, safe, widely applicable approach next-generation

Language: Английский

Citations

2