CRISPR/Cas9-mediated multiplex gene editing of gamma and omega gliadins, paving the way for gliadin-free wheat DOI Creative Commons
Susana Sánchez‐León, Miriam Marín‐Sanz, María H. Guzmán-López

et al.

Journal of Experimental Botany, Journal Year: 2024, Volume and Issue: 75(22), P. 7079 - 7095

Published: Sept. 6, 2024

Wheat is a staple cereal in the human diet. Despite its significance, an increasing percentage of population suffers adverse reactions to wheat, which are triggered by wheat gluten, particularly gliadin fractions. In this study, we employed CRISPR/Cas [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein] multiplexing introduce targeted mutations into γ- and ω-gliadin genes produce lines deficient one or both immunogenic fractions simultaneously. For eight single guide RNAs (sgRNAs) were designed combined four plasmids 59 modified lines, 20 exhibited target genes. Characterization these through Sanger sequencing next-generation revealed complex pattern InDels, including deletions spanning multiple sgRNAs. The transmitted offspring, analysis homozygous derived reverse-phase HPLC monoclonal antibodies showed 97.7% reduction gluten content. Crossing with other α-gliadins allowed be combined. This work represents important step forward use develop gluten-free wheat.

Language: Английский

The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model DOI Creative Commons
Yunfei Li, Junjie Xu, Xuefei Guo

et al.

Genome biology, Journal Year: 2023, Volume and Issue: 24(1)

Published: Feb. 1, 2023

The CRISPR-Cas13 system is an RNA-guided RNA-targeting and has been widely used in transcriptome engineering with potentially important clinical applications. However, it still controversial whether Cas13 exhibits collateral activity mammalian cells.

Language: Английский

Citations

48
Challenges Facing CRISPR/Cas9-Based Genome Editing in Plants DOI Creative Commons
Seungmin Son, Sang Ryeol Park

Frontiers in Plant Science, Journal Year: 2022, Volume and Issue: 13

Published: May 18, 2022

The development of plant varieties with desired traits is imperative to ensure future food security. revolution genome editing technologies based on the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has ushered in a new era breeding. Cas9 and single-guide RNA (sgRNA) form an effective targeting complex locus or loci interest, enabling all plants high accuracy efficiency. Therefore, CRISPR/Cas9 can save both time labor relative what typically associated traditional breeding methods. However, despite improvements gene editing, several challenges remain that limit application CRISPR/Cas9-based plants. Here, we focus four issues relevant editing: (1) organelle editing; (2) transgene-free (3) virus-induced (4) recalcitrant elite crop inbred lines. This review provides up-to-date summary state CRISPR/Cas9-mediated will push this technique forward.

Language: Английский

Citations

69
Genome editing around the globe: An update on policies and perceptions DOI Creative Commons
Thorben Sprink, Ralf Wilhelm, Frank Hartung

et al.

PLANT PHYSIOLOGY, Journal Year: 2022, Volume and Issue: 190(3), P. 1579 - 1587

Published: Aug. 17, 2022

A decade ago, the CRISPR/Cas system has been adapted for genome editing. Since then, hundreds of organisms have altered using editing and discussions were raised on regulatory status edited esp. crops. To date, many countries made decisions products editing, by exempting some kinds edits from classical GMO regulation. However, guidance differs between even in same region. Several are still debating issue or progress updating systems to cover The current global situation different is putting a harmonized framework genome-edited crops far future. In this update, we summarize developments field regulation concerning present short insight into perception society.

Language: Английский

Citations

57
Occurrence and Nature of Off-Target Modifications by CRISPR-Cas Genome Editing in Plants DOI Creative Commons
Mark Sturme, Jan Pieter van der Berg, Lianne M. S. Bouwman

et al.

ACS Agricultural Science & Technology, Journal Year: 2022, Volume and Issue: 2(2), P. 192 - 201

Published: March 3, 2022

CRISPR-Cas-based genome editing allows for precise and targeted genetic modification of plants. Nevertheless, unintended off-target edits can arise that might confer risks when present in gene-edited food crops. Through an extensive literature review we gathered information on CRISPR-Cas Most observed changes were small insertions or deletions (1–22 bp) nucleotide substitutions, large (>100 rare. One study detected the insertion vector-derived DNA sequences, which is important considering risk assessment Off-target sites had few mismatches (1–3 nt) with target sequence mainly located protein-coding regions, often gene homologues. Off-targets predominantly via biased analysis predicted instead unbiased genome-wide analysis. CRISPR-Cas-edited plants showed lower mutation frequencies than conventionally bred This Review aid discussions relevance evaluating modifications

Language: Английский

Citations

52
Genome Editing for Improving Crop Nutrition DOI Creative Commons

Ai Nagamine,

Hiroshi Ezura

Frontiers in Genome Editing, Journal Year: 2022, Volume and Issue: 4

Published: Feb. 9, 2022

Genome editing technologies, including CRISPR/Cas9 and TALEN, are excellent genetic modification techniques being proven to be powerful tools not only in the field of basic science but also crop breeding. Recently, two genome-edited crops targeted for nutritional improvement, high GABA tomatoes oleic acid soybeans, have been released market. Nutritional improvement cultivated has a major target conventional technologies as well classical breeding methods. Mutations created by genome considered almost identical spontaneous mutations because mutation inducer, transformed foreign gene, can completely eliminated from final hosts after causing mutation. Therefore, expected relatively easy supply market, unlike GMO crops. On other hand, due their technical feature, main goal current creation is often total or partial disruption genes rather than gene delivery. obtain desired trait using technology, some cases, different approach that recombination technology may required. In this mini-review, we will review several traits suitable targets editing, examples mentioned above, discuss how an effective improving

Language: Английский

Citations

39
Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review DOI Creative Commons
Fernanda dos Santos Nascimento, Anelita de Jesus Rocha, Julianna Matos da Silva Soares

et al.

Plants, Journal Year: 2023, Volume and Issue: 12(2), P. 305 - 305

Published: Jan. 9, 2023

Agricultural crops are exposed to various abiotic stresses, such as salinity, water deficits, temperature extremes, floods, radiation, and metal toxicity. To overcome these challenges, breeding programs seek improve methods techniques. Gene editing by Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR/Cas-is a versatile tool for in all layers of the central dogma with focus on development cultivars plants resistant or tolerant multiple biotic stresses. This systematic review (SR) brings new contributions study use CRISPR/Cas gene tolerance stress plants. Articles deposited different electronic databases, using search

Language: Английский

Citations

37
Enhancement of specialized metabolites using CRISPR/Cas gene editing technology in medicinal plants DOI Creative Commons
Swati Das, Moonhyuk Kwon, Jae‐Yean Kim

et al.

Frontiers in Plant Science, Journal Year: 2024, Volume and Issue: 15

Published: Feb. 21, 2024

Plants are the richest source of specialized metabolites. The metabolites offer a variety physiological benefits and many adaptive evolutionary advantages frequently linked to plant defense mechanisms. Medicinal plants vital nutrition active pharmaceutical agents. production valuable bioactive compounds has increased with improvement transgenic techniques like gene silencing overexpression. These beneficial for decreasing costs increasing nutritional value. Utilizing biotechnological applications enhance in medicinal needs characterization identification genes within an elucidated pathway. breakthrough advancement CRISPR/Cas-based editing improving specific have gained significant importance contemporary times. This article imparts comprehensive recapitulation latest advancements made implementation CRISPR-gene purpose augmenting plants. We also provide further insights perspectives metabolic engineering scenarios

Language: Английский

Citations

15
Emerging Gene Therapeutics for Epidermolysis Bullosa under Development DOI Open Access
Johannes Bischof, Markus Hierl, Ulrich Koller

et al.

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

Citations

14
Crop Improvement: Comparison of Transgenesis and Gene Editing DOI Creative Commons
Natalya V. Permyakova, Е. В. Дейнеко

Horticulturae, Journal Year: 2024, Volume and Issue: 10(1), P. 57 - 57

Published: Jan. 6, 2024

The development and improvement of molecular biology methods have led to the creation new technologies that make it possible modify plant genomes by transferring integrating into genomes’ heterologous genes from various expression systems (genetic engineering), as well inducing knockouts one or more target interest (genomic editing). genome-editing is a milestone in modern breeding certainly relies on knowledge developed for transgenesis. This review will discuss issues related advantages disadvantages both improving economically valuable traits important crops.

Language: Английский

Citations

12
Cellular engineering of plant cells for improved therapeutic protein production DOI Open Access

Uddhab Karki,

Hong Fang, Wenzheng Guo

et al.

Plant Cell Reports, Journal Year: 2021, Volume and Issue: 40(7), P. 1087 - 1099

Published: April 10, 2021

Language: Английский

Citations

47