CRISPR/Cas9‐Based Genome Editing and its Applications for Functional Genomic Analyses in Plants

Li, Jun; Li, Yán; Ma, Ligeng

doi:10.1002/smtd.201800473

Cited by 28 publications

(13 citation statements)

References 205 publications

(376 reference statements)

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“…Several genome editing technologies with the ability to change the code of life with high specificity have been developed recently (Li et al 2019 ). Advances in genome editing have revolutionized life science, including plant science.…”

Section: Introductionmentioning

confidence: 99%

Recent advances in CRISPR/Cas9 and applications for wheat functional genomics and breeding

2021

aBIOTECH

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Common wheat (Triticum aestivum L.) is one of the three major food crops in the world; thus, wheat breeding programs are important for world food security. Characterizing the genes that control important agronomic traits and finding new ways to alter them are necessary to improve wheat breeding. Functional genomics and breeding in polyploid wheat has been greatly accelerated by the advent of several powerful tools, especially CRISPR/Cas9 genome editing technology, which allows multiplex genome engineering. Here, we describe the development of CRISPR/Cas9, which has revolutionized the field of genome editing. In addition, we emphasize technological breakthroughs (e.g., base editing and prime editing) based on CRISPR/Cas9. We also summarize recent applications and advances in the functional annotation and breeding of wheat, and we introduce the production of CRISPR-edited DNA-free wheat. Combined with other achievements, CRISPR and CRISPR-based genome editing will speed progress in wheat biology and promote sustainable agriculture.

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Section: Introductionmentioning

confidence: 99%

Recent advances in CRISPR/Cas9 and applications for wheat functional genomics and breeding

2021

aBIOTECH

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“…Two major components are essential for typical engineered CRISPR-Cas systems, a Cas endonuclease protein, and a single guide RNA (sgRNA) of 20 nucleotide sequences that guide the Cas enzyme to the target sequence for introducing the double-stranded break (DSB) ( Mahfouz et al, 2014 ). Multiple variants of Cas9 and gRNA are available according to their novel application in the field of genetic engineering in plants ( Chen et al, 2019 ; Li et al, 2019 ).…”

Section: Crispr-based Genome Editing Tool: Components and Mechanismmentioning

confidence: 99%

New Hope for Genome Editing in Cultivated Grasses: CRISPR Variants and Application

et al. 2022

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With the advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas) mediated genome editing, crop improvement has progressed significantly in recent years. In this genome editing tool, CRISPR-associated Cas nucleases are restricted to their target of DNA by their preferred protospacer adjacent motifs (PAMs). A number of CRISPR-Cas variants have been developed e.g. CRISPR-Cas9, -Cas12a and -Cas12b, with different PAM requirements. In this mini-review, we briefly explain the components of the CRISPR-based genome editing tool for crop improvement. Moreover, we intend to highlight the information on the latest development and breakthrough in CRISPR technology, with a focus on a comparison of major variants (CRISPR-Cas9, -Cas12a, and -Cas12b) to the newly developed CRISPR-SpRY that have nearly PAM-less genome editing ability. Additionally, we briefly explain the application of CRISPR technology in the improvement of cultivated grasses with regard to biotic and abiotic stress tolerance as well as improving the quality and yield.

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“…There has been a remodeling of the Cas protein, specifically the protein designated SpCas9 or simply Cas9, which is from Streptococcus pyogenes. Generation of these protein variants increased options beyond the ability to induce double strand breaks to now having a variant that cleaves only one strand, referred to as a Cas9 nickase, and a variant that binds DNA, but does not cleave, referred to as a "dead" Cas9 (dCas9) (19). These variants have been exploited for development of new designs that make it possible to achieve modifications that range from single base pair changes to exchanging of alleles [20,21].…”

Section: Expanding the Crispr/cas Toolboxmentioning

confidence: 99%

“…The initially available and subsequently most widely used Cas protein, Cas9, results in blunt-end cuts of the DNA a few base pairs upstream from the recognized PAM sequence, 5'-NGG-3' (N = A, T, G, C) (19). Cas9 has been shown to be effective in editing genomes of numerous organisms including plants with some groups developing plant codon optimized versions depending on the species of interest.…”

Section: The Cas Protein Collectionmentioning

confidence: 99%

“…The number of genomes modified by the creation of indels, including some important food crops such as maize (13), potato (14), rice [14], tomato (15), and wheat (16), has grown enormously since the very first reports of the application of CRISPR-mediated editing in plants. There are many reviews that provide information related to modified plant genomes with CRISPR/Cas-mediated editing in addition to the traits of interest (17)(18)(19).…”

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Applying gene editing to tailor precise genetic modifications in plants

Eck

2020

Journal of Biological Chemistry

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The ability to tailor alterations in genomes, including plant genomes, in a site-specific manner has been greatly advanced through approaches that reduced the complexity and time of genome sequencing along with development of gene editing technologies. These technologies provide a valuable foundation for studies of gene function, metabolic engineering and trait modification for crop improvement. Development of genome editing methodologies began approximately 20 years ago first with meganucleases followed by Zinc Finger Nucleases, Transcriptional Activator-Like Effector Nucleases and, most recently, Clustered Regulatory Interspaced Short Palindromic Repeat (CRISPR)-Associated Protein (Cas) (CRISPR/Cas), which is by far the most utilized method. The premise of CRISPR/Cas centers on the cleaving of one or both DNA strands by a Cas protein, an endonuclease, followed by mending of the DNA by repair mechanisms inherent in cells. Its user-friendly construct design, greater flexibility in targeting genomic regions, and the cost-effective attributes have resulted in it being widely adopted and revolutionizing precise modification of the genomes of many organisms. Indeed, the CRISPR/Cas system has been utilized for gene editing in many plant species including important food crops such as maize, wheat, rice, and potatoes. This review summarizes the various approaches including the most recent designs being used to make modifications from as small as a single base pair change to insertion of DNA fragments. On the gene expression level, strategies are presented that make it possible to knock-out or modulate through activation and repression. Also discussed are pre-requisites necessary for CRISPR/Cas-mediated editing as well as the current challenges.

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CRISPR/Cas9‐Based Genome Editing and its Applications for Functional Genomic Analyses in Plants

Cited by 28 publications

References 205 publications

Recent advances in CRISPR/Cas9 and applications for wheat functional genomics and breeding

Recent advances in CRISPR/Cas9 and applications for wheat functional genomics and breeding

New Hope for Genome Editing in Cultivated Grasses: CRISPR Variants and Application

Applying gene editing to tailor precise genetic modifications in plants

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