CRISPR-based genome editing in wheat: a comprehensive review and future prospects

Kumar, Rakesh; Kaur, Amandeep; Pandey, Ankita; Mamrutha, H. M.; Singh, Gyanendra

doi:10.1007/s11033-019-04761-3

Cited by 52 publications

(41 citation statements)

References 82 publications

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“…These repairs can be tricked to add, remove, or substitute a series of nucleotides in the genetic code, thus enabling the introduction of known desired alleles in the target organism [22]. The major DNA DSB repair mechanisms include Non-Homologous End Joining (NHEJ) and Homologous Recombination (HR) [19,23]. In the NHEJ repair pathway, the two broken ends are joined with a loss of few nucleotides hence it is a mutation prone process and taken advantage in knocking out of gene function in gene editing [24].…”

Section: What Is the Gene-editing Technique?mentioning

confidence: 99%

“…HR is a highly precise repair mechanism and exploited in genome editing techniques to edit single nucleotides or to introduce other precise changes in the genome [25]. Palindromic Repeats (CRISPR/Cas9) system [19,26]. Among all these, CRISPR/cas9 is the latest gene-editing tool that is highly accurate, rapid, simple and comparatively cheaper than other gene-editing tools [27,28].…”

Section: What Is the Gene-editing Technique?mentioning

confidence: 99%

“…Cas9 (CRISPR-associated protein 9) was adopted from the defence machinery of bacteria [19,25,28]. When a virus (Bacteriophage) attacks a bacteria, the bacteria captures snippets of the genetic material of the virus (DNA) and synthesize DNA segments that are known as CRISPR arrays [19,25,26]. These CRISPR arrays memorize the virus, on future invasions of the same or similar virus, the bacteria synthesize the RNA segments from the CRISPR arrays that target the virus.…”

Section: What Is Crispr-cas9?mentioning

confidence: 99%

“…In recent years, gene editing techniques (ZFN, TALEN) emerged as a promising approach to edit or delete the toxic gluten fractions in wheat [19]. CRISPR/Cas9 is a novel secondgeneration genome editing system.…”

Section: Introductionmentioning

confidence: 99%

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Status and Contribution of CRISPR/Cas9 Based Gene-Editing System in the Development of a Low-Immunogenic Wheat Variety

Verma¹,

Tiwari²,

Lionetti³

et al. 2020

Preprint

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Gluten, a wheat protein, contains epitopes that trigger celiac disease (CD). So far, there is no treatment available for CD other than following a life-long, strict gluten-free diet (GFD). A very low-gluten or gluten-free wheat could provide an alternative treatment to CD. Till date, conventional plant breeding methods are not sufficient to produce celiac-safe wheat. Clustered Regularly Interspaced Short Palindromic Repeats-associated protein 9 (CRISPR/Cas9) is a versatile gene-editing system may efficiently edit the immunogenic gluten protein thereby producing a celiac-safe wheat variety. However, CRISPR/Cas9-mediated genome editing system has not been widely investigated to edit the wheat genome. Published literature available on various scientific platforms, that applied the CRISPR/Cas9 system to edit the wheat genome were explored. Only original research articles were included. Review articles, protocols, scientific presentations, and Ph.D. thesis were excluded. CRISPR/Cas9 is a highly specific gene-editing technology that can be used to efficiently edit the complex hexaploid wheat genome. It targets a specific wheat genome locus, delete an immunogenic gene and replace it with a preferred gene. CRISPR/Cas9 technology could be a breakthrough in providing an alternative treatment for CD. However, further studies are required to efficiently apply this gene-editing technology to develop a celiac-safe wheat variety.

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Section: What Is the Gene-editing Technique?mentioning

confidence: 99%

Section: What Is the Gene-editing Technique?mentioning

confidence: 99%

Section: What Is Crispr-cas9?mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Status and Contribution of CRISPR/Cas9 Based Gene-Editing System in the Development of a Low-Immunogenic Wheat Variety

Verma¹,

Tiwari²,

Lionetti³

et al. 2020

Preprint

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“…This is further limited by the requirement for careful site selection to minimize off-target binding and cleavage based on the tolerance for mismatches in the gRNA-PAM-target complex [19][20][21][22][23] . This constraint becomes particularly evident in therapeutic applications where even rare genome alterations resulting from offtargets are undesirable or when targeting more structurally complex plant genomes 24,25 . Moreover, these restraints impact the use of Cas9 for homology-directed repair (HDR), template-free editing, base editing, or prime editing applications, where the outcome is reliant on the proximity of the desired change to the target sequence [26][27][28][29][30][31][32] .…”

Section: Introductionmentioning

confidence: 99%

Biochemically diverse CRISPR-Cas9 orthologs

Gasiūnas

Young

Karvelis

et al. 2020

Preprint

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CRISPR-Cas9 nucleases are abundant in microbes. To explore this largely uncharacterized diversity, we applied cell-free biochemical screens to rapidly assess the protospacer adjacent motif (PAM) and guide RNA (gRNA) requirements of novel Cas9 proteins. This approach permitted the characterization of 79Cas9 orthologs with at least 7 distinct classes of gRNAs and 50 different PAM sequence requirements.PAM recognition spanned the entire spectrum of T-, A-, C-, and G-rich nucleotides ranging from simple di-nucleotide recognition to complex sequence strings longer than 4. Computational analyses indicated that most of this diversity came from 4 groups of interrelated sequences providing new insight into Cas9 evolution and efforts to engineer PAM recognition. A subset of Cas9 orthologs were purified and their activities examined further exposing additional biochemical diversity. This constituted both narrow and broad ranges of temperature dependence, staggered-end DNA target cleavage, and a requirement for longer stretches of homology between gRNA and DNA target to function robustly. In all, the diverse collection of Cas9 orthologs presented here sheds light on Cas9 evolution and provides a rich source of PAM recognition and other potentially desirable properties that may be mined to expand the genome editing toolbox with new RNA-programmable nucleases.

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