Application of CRISPR/Cas-mediated base editing for directed protein evolution in plants

Capdeville, Niklas; Schindele, Patrick; Puchta, Holger

doi:10.1007/s11427-020-1655-9

Cited by 5 publications

(6 citation statements)

References 19 publications

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“…CRISPR-Cas9 has been successfully exploited for directed protein evolution in the native plant environment by coupling Cas9 with a library of sgRNAs tilling all potential sites on both strands of the relevant coding sequence (Butt et al, 2019). Nevertheless, the predominant mutations induced by Cas9 are indel-associated frameshifts, making it difficult to generate all the amino acid substitutions needed for the generation of SNPs, the most common sources of genetic variation in nature (Capdeville et al, 2020). Base editors are tools ideally suited for overcoming this difficulty.…”

Section: Haploid Induction and Artificial Apomixismentioning

confidence: 99%

Genome engineering for crop improvement and future agriculture

Gao

2021

Cell

513

340

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Section: Haploid Induction and Artificial Apomixismentioning

confidence: 99%

Genome engineering for crop improvement and future agriculture

Gao

2021

Cell

513

340

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“…A different technology has been established that enables DSB and donor template-free base editing from C to T at a specific target site [27]. ese base substitutions are mediated by a cytidine deaminase that converts cytidine into uridine by eliminating an amino group.…”

Section: Crispr/cas9 Technology In Plant Gementioning

confidence: 99%

“…ese base substitutions are mediated by a cytidine deaminase that converts cytidine into uridine by eliminating an amino group. Consequent DNA repair of the U-G mismatch results in a U-A base pair that is further resolved to a stable T-A base pair [27]. CRISPR/Casbased directed evolution is a developing field, in which base editing plays a key role, and in recent years, numerous groups have established methods to improve base editing [27][28][29].…”

Section: Crispr/cas9 Technology In Plant Gementioning

confidence: 99%

“…Consequent DNA repair of the U-G mismatch results in a U-A base pair that is further resolved to a stable T-A base pair [27]. CRISPR/Casbased directed evolution is a developing field, in which base editing plays a key role, and in recent years, numerous groups have established methods to improve base editing [27][28][29]. e viability and effectiveness of cytidine deaminase base editing (CBE) have been tested in rice at three chosen targets: one target (P2) in OsPDS, which encodes a phytoene desaturase, and two targets (S3 and S5) in OsS-BEIIb, which encode starch branching enzyme IIb.…”

Section: Crispr/cas9 Technology In Plant Gementioning

confidence: 99%

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The Application of CRISPR/Cas9 Technology in the Management of Genetic and Nongenetic Plant Traits

Mutezo

Sedibe

Mofokeng

et al. 2021

International Journal of Agronomy

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Gene editing (GE) has yielded positive results in the management of genetic and nongenetic plant traits. Clustered interspaced palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) (together known as the CRISPR/Cas system) are relatively easy to use, compared to other existing genome editing technologies. The CRISPR/Cas tool produces unchanging gene mutations, with the ability to segregate from the Cas9/sgRNA construct to avoid similar modifications by CRISPR/Cas. CRISPR/Cas GE is fast and accurate; crops developed using this technique are resistant to viruses, fungi, and bacteria and resilient to abiotic and biotic stresses, while crops established using conventional methods take between 10 and 15 years to develop resistance. Therefore, CRISPR/Cas is a useful tool for sustainable agricultural production. Plant traits have been successfully manipulated using this technology. Notwithstanding the technical challenges of transferring CRISPR/Cas9 developed crops from the laboratory to the field, additional obstacles include uncertain policy systems, dispute over intellectual property ownership, and acceptability by consumers. Several Cas9-based applied techniques have gained popularity that enable researchers to enhance plants with speed and accuracy. In conclusion, the CRISPR/Cas9 system is a powerful technology for genetically modifying crops.

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“…Since the CRISPR‐Cas system can create targeted mutagenesis in situ , the CRISPR‐based directed evolution has become a powerful plant breeding biotechnology to improve agronomic traits in plants (Butt et al ., 2019; Kuang et al ., 2020; Li et al ., 2020a; Liu et al ., 2020; Wang et al ., 2022; Xu et al ., 2021b). In particular, the DNA double‐strand break (DSB)‐independent genome editing tools, such as base editors and prime editors, are deemed suitable for performing saturated targeted endogenous mutagenesis in plants via introducing base substitutions (Capdeville et al ., 2020; Xu et al ., 2021b).…”

Section: Introductionmentioning

confidence: 99%

Directed evolution rice genes with randomly multiplexed sgRNAs assembly of base editors

Zhang

Shan

Sun

et al. 2023

Plant Biotechnology Journal

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SummaryCRISPR‐based directed evolution is an effective breeding biotechnology to improve agronomic traits in plants. However, its gene diversification is still limited using individual single guide RNA. We described here a multiplexed orthogonal base editor (MoBE), and a randomly multiplexed sgRNAs assembly strategy to maximize gene diversification. MoBE could induce efficiently orthogonal ABE (<36.6%), CBE (<36.0%), and A&CBE (<37.6%) on different targets, while the sgRNA assembling strategy randomized base editing events on various targets. With respective 130 and 84 targets from each strand of the 34th exon of rice acetyl‐coenzyme A carboxylase (OsACC), we observed the target‐scaffold combination types up to 27 294 in randomly dual and randomly triple sgRNA libraries. We further performed directed evolution of OsACC using MoBE and randomly dual sgRNA libraries in rice, and obtained single or linked mutations of stronger herbicide resistance. These strategies are useful for in situ directed evolution of functional genes and may accelerate trait improvement in rice.

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Application of CRISPR/Cas-mediated base editing for directed protein evolution in plants

Cited by 5 publications

References 19 publications

Genome engineering for crop improvement and future agriculture

Genome engineering for crop improvement and future agriculture

The Application of CRISPR/Cas9 Technology in the Management of Genetic and Nongenetic Plant Traits

Directed evolution rice genes with randomly multiplexed sgRNAs assembly of base editors

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