Optimizing plant adenine base editor systems by modifying the transgene selection system

Li, Juan; Qin, Ruiying; Zhang, Yuandi; Xu, Shanbin; Liu, Xiaoshuang; Yang, Jian‐Bo; Zhang, Xiuqing; Wei, Pengcheng

doi:10.1111/pbi.13304

Cited by 18 publications

(8 citation statements)

References 11 publications

(12 reference statements)

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“…Tang and colleagues presented data from rice protoplasts only and did not obtain any stable transgenic lines with editing events (Tang et al , 2020), whereas 1‐3 rice endogenous genes were edited with relatively low efficiencies in transgenic plants in the other studies. It is possible that the editing efficiency of the prime editing system can be improved by using other reverse transcriptases (Stamos et al , 2017) or modifying the transgene selection system (Li et al , 2019) to make the prime editors powerful tools for precision molecular breeding of crops.…”

Section: Figurementioning

confidence: 99%

Precision genome engineering in rice using prime editing system

Hua

Jiang

Tao

et al. 2020

Plant Biotechnology Journal

131

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Section: Figurementioning

confidence: 99%

Precision genome engineering in rice using prime editing system

Hua

Jiang

Tao

et al. 2020

Plant Biotechnology Journal

131

View full text Add to dashboard Cite

“…To expand the editing scope, we replaced the Sp Cas9 in the base editors with Cas orthologs or evolved Sp Cas9 variants with relaxed protospacer adjacent motif (PAM) recognition ( Hua et al., 2019a , Hua et al., 2019b , Qin et al., 2019a , Ren et al., 2019 , Zhong et al., 2019 , Wang et al., 2020 ). Furthermore, the efficiency of base editors was enhanced by modification of fusion protein structure, utilization of deaminase orthologs, or development of selection-enriched systems ( Li et al., 2018 , Li et al., 2020c , Zong et al., 2018 , Qin et al., 2019b , Hua et al., 2020 , Xu et al., 2020a ). Editing by base editors has outperformed HDR-mediated substitutions in plants in terms of efficiency and product purity.…”

Section: Introductionmentioning

confidence: 99%

Development of Plant Prime-Editing Systems for Precise Genome Editing

Liu

et al. 2020

Plant Communications

Self Cite

160

123

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Prime-editing systems have the capability to perform efficient and precise genome editing in human cells. In this study, we first developed a plant prime editor 2 (pPE2) system and test its activity by generating a targeted mutation on an HPT -ATG reporter in rice. Our results showed that the pPE2 system could induce programmable editing at different genome sites. In transgenic T 0 plants, pPE2-generated mutants occurred with 0%–31.3% frequency, suggesting that the efficiency of pPE2 varied greatly at different genomic sites and with prime-editing guide RNAs of diverse structures. To optimize editing efficiency, guide RNAs were introduced into the pPE2 system following the PE3 and PE3b strategy in human cells. However, at the genomic sites tested in this study, pPE3 systems generated only comparable or even lower editing frequencies. Furthemore, we developed a surrogate pPE2 system by incorporating the HPT -ATG reporter to enrich the prime-edited cells. The nucleotide editing was easily detected in the resistant calli transformed with the surrogate pPE2 system, presumably due to the enhanced screening efficiency of edited cells. Taken together, our results indicate that plant prime-editing systems we developed could provide versatile and flexible editing in rice genome.

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“…For an approach requiring a high level of outcome predictability, UGI domain(s) can be added to the CBE architecture, resulting in a higher rate of C-to-T substitutions with lower unwanted mutations ( Zong et al., 2017 ; Qin et al., 2019b ). As observed in animals, ABEs produce A-to-G transitions in plants, with a very low rate of indels ( Li et al., 2018a , 2019b ; Kang et al., 2018 ; Yan et al., 2018 ; Hao et al., 2019 ; Negishi et al., 2019 ; Hua et al., 2020b ). These BEs allowed the production of plants with new agronomic traits, including pathogen resistance ( Bharat et al., 2020 ; Mishra et al., 2020 ).…”

Section: Precision Editing: Refining the Tools?mentioning

confidence: 67%

Precision Breeding Made Real with CRISPR: Illustration through Genetic Resistance to Pathogens

Veillet

Durand

Kroj

et al. 2020

Plant Communications

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Since its discovery as a bacterial adaptive immune system and its development for genome editing in eukaryotes, the CRISPR technology has revolutionized plant research and precision crop breeding. The CRISPR toolbox holds great promise in the production of crops with genetic disease resistance to increase agriculture resilience and reduce chemical crop protection with a strong impact on the environment and public health. In this review, we provide an extensive overview on recent breakthroughs in CRISPR technology, including the newly developed prime editing system that allows precision gene editing in plants. We present how each CRISPR tool can be selected for optimal use in accordance with its specific strengths and limitations, and illustrate how the CRISPR toolbox can foster the development of genetically pathogen-resistant crops for sustainable agriculture.

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Optimizing plant adenine base editor systems by modifying the transgene selection system

Cited by 18 publications

References 11 publications

Precision genome engineering in rice using prime editing system

Precision genome engineering in rice using prime editing system

Development of Plant Prime-Editing Systems for Precise Genome Editing

Precision Breeding Made Real with CRISPR: Illustration through Genetic Resistance to Pathogens

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