Shortening the sgRNA-DNA interface enables SpCas9 and eSpCas9(1.1) to nick the target DNA strand

Fan, Rong; Chai, Zhuangzhuang; Xing, Sinian; Chen, Kunling; Qiu, Fengti; Chai, Tuanyao; Qiu, Jian; Zhengbin, Zhang; Zhang, Huawei; Gao, Caixia

doi:10.1007/s11427-020-1722-0

Cited by 10 publications

(4 citation statements)

References 30 publications

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“…Cytosine base editing uses cytidine deaminase and Cas9 fusion proteins to alter nucleotides in target DNA in situ, generating C:G>T:A base transitions (Komor et al, 2016;Xue et al, 2018). So far point mutations have been introduced into several genes, including acetolactate synthase (ALS), acetyl-coenzyme A carboxylase and tubulin genes, to generate herbicide-tolerant plants (Fan et al, 2020;Li et al, 2018;Liu et al, 2020a;Liu et al, 2020b;Zhang et al, 2019). ALS encodes a key enzyme in the biosynthesis of branched-chain amino acids, which is the target of a large number of herbicides with dissimilar chemistries, including sulfonylurea (SU), imidazolinone (IMI), triazolopyrimidine (TP), pyrimidinyl-thiobenzoates (PTB) and sulfonyl-aminocarbonyltriazolinone (SCT).…”

Section: Introductionmentioning

confidence: 99%

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

Zhang

Chen

Meng

et al. 2020

Sci. China Life Sci.

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Herbicide-tolerant rice varieties generated by genome editing are highly desirable for weed control. We have used a cytosine base editor to create a series of missense mutations in the P171 and/or G628 codons of the acetolactate synthase (ALS) gene to confer herbicide tolerance in rice. The four different missense mutations in the P171 codon, P171S, P171A, P171Y and P171F, exhibited different patterns of tolerance towards five representative herbicides from five chemical families of ALS inhibitors. For example, P171S and P171A had lower levels of tolerance than P171Y and P171F to bispyribac but not to the other herbicides. Interestingly, a novel triple mutant (P171F/G628E/G629S) had the highest tolerance to all five tested herbicides. Field trials showed that both P171F and P171F/G628E/G629S could potentially be used with nicosulfuron. Our work illustrates an effective way of using base editing to generate herbicide tolerance in elite rice varieties. base editing, herbicide tolerance, rice, acetolactate synthase (ALS)

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Section: Introductionmentioning

confidence: 99%

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

Zhang

Chen

Meng

et al. 2020

Sci. China Life Sci.

Self Cite

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“…In addition, the introduction of a pair of singleguide RNAs enables nCas9 to insert a third style of edit, known as indels. Another research used a dual-function framework consisting of a truncated protospacer and a full-length protospacer to regulate the function of an improved specificity SpCas9 variant 1.1-based CBE to induce an indel and C:G→T:A base alterations in plants [149]. Multiplexed orthogonal genome editing in rice was achieved by combining the CBE and ABE based on SpCas9 and SaCas9, respectively [20,150].…”

Section: Multiplex Gene Editingmentioning

confidence: 99%

CRISPR-Cas Mediated Genome Editing: A Paradigm Shift towards Sustainable Agriculture and Biotechnology

Rehman¹,

Zafar²,

Ali³

et al. 2022

APRJ

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CRISPR–Cas genome editing technology developed from prokaryotes has transformed the molecular biology of plants past all assumptions. CRISPR–Cas, which is distinguished by its resilience, relatively high specificity, and easy implementation, enables specific genetic modification of crops, allowing for the creation of germplasms with favorable characters and the development of innovative, highly efficient agricultural systems. Moreover, many new biotechnologies in the framework of CRISPR–Cas platforms have bolstered basic research as well as synthetic biology toolkit of plants. In this article, initially, we provide a brief overview of CRISPR–Cas gene editing, emphasis on the modern, most specific gene-editing techniques, such as prime and base editing. Following that, the major role of CRISPR–Cas in plants in enhancing pesticide and disease resistance, quality, yield, breeding, and faster domestication are next discussed. In this review, we discuss the current advancements in plant biotechnology linked to CRISPR–Cas, such as CRISPR–Cas gene control, reagent conveyance, multiplexed gene editing, directed evolution, and mutagenesis. In the end, we talk about how this innovative technology may be used in the future.

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“…However, due to its natural off-target effects, researchers are making various attempts in this area to improve the situation. Researchers have identified variants of Cas9, such as SpCas9HF1, eSpCas9, and HypaCas, [103][104][105] with reduced offtarget effects. Future research should focus on minimizing off-target effects when performing high-fidelity nucleic acid detection.…”

Section: Discussionmentioning

confidence: 99%

Cas-based bacterial detection: recent advances and perspectives

Lan,

Shu,

Jiang

et al. 2024

Analyst

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Shortening the sgRNA-DNA interface enables SpCas9 and eSpCas9(1.1) to nick the target DNA strand

Cited by 10 publications

References 30 publications

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

CRISPR-Cas Mediated Genome Editing: A Paradigm Shift towards Sustainable Agriculture and Biotechnology

Cas-based bacterial detection: recent advances and perspectives

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