CRISPR/dCas9-Based Systems: Mechanisms and Applications in Plant Sciences

Karlson, Chou Khai Soong; Noor, Siti Nurfadhlina Mohd; Nolte, Nadja; Tan, Boon Chin

doi:10.3390/plants10102055

Cited by 47 publications

(35 citation statements)

References 162 publications

(180 reference statements)

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“…Although there are several CRISPR/Cas technologies based on different Cas activities [ 104 , 108 ], the underlying molecular mechanism is the same in all cases. The two main components of such systems are the gRNA and the Cas nuclease.…”

Section: Editing the Plant Genome In Transgenic Hairy Roots: Vector Componentsmentioning

confidence: 99%

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Kiryushkin

Ilina

Guseva

et al. 2021

Plants

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CRISPR/Cas-mediated genome editing is a powerful tool of plant functional genomics. Hairy root transformation is a rapid and convenient approach for obtaining transgenic roots. When combined, these techniques represent a fast and effective means of studying gene function. In this review, we outline the current state of the art reached by the combination of these approaches over seven years. Additionally, we discuss the origins of different Agrobacterium rhizogenes strains that are widely used for hairy root transformation; the components of CRISPR/Cas vectors, such as the promoters that drive Cas or gRNA expression, the types of Cas nuclease, and selectable and screenable markers; and the application of CRISPR/Cas genome editing in hairy roots. The modification of the already known vector pKSE401 with the addition of the rice translational enhancer OsMac3 and the gene encoding the fluorescent protein DsRed1 is also described.

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Section: Editing the Plant Genome In Transgenic Hairy Roots: Vector Componentsmentioning

confidence: 99%

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Kiryushkin

Ilina

Guseva

et al. 2021

Plants

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“…However, their mutation ratios are low, and the mutation sites are random and unknown. The CRISPR/Cas9 system can easily produce target gene mutation, and has been widely used in plant mutant creation, especially for reverse genetics in revealing target gene function and regulation [6][7][8][9][10]. The Cas9 protein functions as a nuclease and is guided to the target site by the engineered single guide RNA (gRNA) including 20 specific nucleotides of the selected gene to determine the site-specific targeting.…”

Section: Discussionmentioning

confidence: 99%

“…The reverse genetics can reveal the gene function through the mutation or expression regulation of target gene [1,2]. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) system can easily mutate and edit the target gene, and has been widely used in plant sciences and crop improvement [6][7][8][9][10]. The mutants induced by CRISPR/Cas system usually have many heterozygous and bi-allelic mutant lines, which can produce the homozygous mutant plants in their offspring segregation population [11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Jiang

Ren

Wang

et al. 2022

Plants

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Bi-allelic mutant lines induced by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) systems are important genetic materials. It is very important to establish a rapid and cheap method in identifying homozygous mutant plants from offspring segregation populations of bi-allelic mutant lines. In this study, the offspring genotypes of rice bi-allelic starch branching enzyme IIb mutant lines were identified using the allele specific PCR (AS-PCR) method. The target sequences of two alleles were aligned from their 5′ to 3′ ends, and the first different bases were used as the 3′ ends of mismatch primers. Another mismatched base was introduced at the third nucleotide from the 3′ end of mismatch primer. The PCR reaction mixture and amplification program were optimized according to the differences of mutation target sequence and mismatch primers. The offspring plant genotypes of bi-allelic mutant lines could be accurately identified using the amplified DNA fragments by agarose gel electrophoresis. This study could provide a method reference for the rapid screening of homozygous mutant plants from offspring segregation population of heterozygous and bi-allelic mutant lines.

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“…Therefore, deciphering the epigenetic factors contributing to the pre-harvest sprouting resistance regulation in rice will lay a foundation for a deep understanding of the full machinery in real environmental cues. Moreover, an intensive validation of the existing candidate genes should be processed via CRISPR-Cas9 [ 129 ], RNAi [ 130 ], and super- Agrobacterium tumefaciens -mediated transformation [ 131 ] as a notable example. Altogether, post-transcriptional regulation encompassing, splicing RNA, and epigenetics offer novel avenues for unravelling the mechanism of resistance of pre-harvest sprouting in rice.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Seed Dormancy and Pre-Harvest Sprouting in Rice—An Updated Overview

Sohn

Subramanian

Thamilarasan

et al. 2021

IJMS

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Pre-harvest sprouting is a critical phenomenon involving the germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. As it results in reduced grain yield and quality, it is a common problem for the farmers who have cultivated the rice and wheat across the globe. Crop yields need to be steadily increased to improve the people’s ability to adapt to risks as the world’s population grows and natural disasters become more frequent. To improve the quality of grain and to avoid pre-harvest sprouting, a clear understanding of the crops should be known with the use of molecular omics approaches. Meanwhile, pre-harvest sprouting is a complicated phenomenon, especially in rice, and physiological, hormonal, and genetic changes should be monitored, which can be modified by high-throughput metabolic engineering techniques. The integration of these data allows the creation of tailored breeding lines suitable for various demands and regions, and it is crucial for increasing the crop yields and economic benefits. In this review, we have provided an overview of seed dormancy and its regulation, the major causes of pre-harvest sprouting, and also unraveled the novel avenues to battle pre-harvest sprouting in cereals with special reference to rice using genomics and transcriptomic approaches.

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CRISPR/dCas9-Based Systems: Mechanisms and Applications in Plant Sciences

Cited by 47 publications

References 162 publications

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Seed Dormancy and Pre-Harvest Sprouting in Rice—An Updated Overview

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