A straightforward plant prime editing system enabled highly efficient precise editing of rice Waxy gene

Li, Zhongsen; Ma, Rui; Liú, Dan; Wang, Ming yue; Zhu, Ting; Deng, Yanxue

doi:10.1016/j.plantsci.2022.111400

Cited by 12 publications

(4 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Subsequently, they were the first to introduce paired pegRNAs and developed a pegRNA designer webpage for facilitating paired pegRNAs selection (Lin et al ., 2021 ). Though not all the strategies helped to improve PE performance, the pioneering PE works in plants have inspired many plant groups to study and apply PE for precision plant breeding in monocots (Butt et al ., 2020 ; Hua et al ., 2020 ; Jiang et al ., 2020 , 2022b ; Li et al ., 2020a , 2022a , 2022b , 2022e , 2023b ; Liang et al ., 2023 ; Lin et al ., 2020 , 2021 ; Molla et al ., 2021 ; Ni et al ., 2023 ; Qiao et al ., 2023 ; Tang et al ., 2020 ; Wang et al ., 2021a ; Xu et al ., 2020a , 2021b , 2022 ; Zou et al ., 2022 ), dicots (Biswas et al ., 2022 ; Jiang et al ., 2022c ; Lu et al ., 2021 ; Perroud et al ., 2022 ; Vu et al ., 2022b ; Wang et al ., 2021a ) and moss ( Physcomitrella patens) (Perroud et al ., 2022 , 2023 ) (Table 1 ). Recently, using the PE2max and pU6 composite promoter driving epegRNAs expression (Figure 2c ), PE efficiency reached up to 40% in rice, 21.5% in maize and 74.5% in wheat, which are sufficiently high for practical applications in plant breeding (Li et al ., 2022b ; Ni et al ., 2023 ; Qiao et al ., 2023 ).…”

Section: Pe Applications In Plantsmentioning

confidence: 99%

Prime editing: Mechanism insight and recent applications in plants

Vu,

Nguyen,

Kim

et al. 2023

Plant Biotechnology Journal

View full text Add to dashboard Cite

SummaryPrime editing (PE) technology utilizes an extended prime editing guide RNA (pegRNA) to direct a fusion peptide consisting of nCas9 (H840) and reverse transcriptase (RT) to a specific location in the genome. This enables the installation of base changes at the targeted site using the extended portion of the pegRNA through RT activity. The resulting product of the RT reaction forms a 3′ flap, which can be incorporated into the genomic site through a series of biochemical steps involving DNA repair and synthesis pathways. PE has demonstrated its effectiveness in achieving almost all forms of precise gene editing, such as base conversions (all types), DNA sequence insertions and deletions, chromosomal translocation and inversion and long DNA sequence insertion at safe harbour sites within the genome. In plant science, PE could serve as a groundbreaking tool for precise gene editing, allowing the creation of desired alleles to improve crop varieties. Nevertheless, its application has encountered limitations due to efficiency constraints, particularly in dicotyledonous plants. In this review, we discuss the step‐by‐step mechanism of PE, shedding light on the critical aspects of each step while suggesting possible solutions to enhance its efficiency. Additionally, we present an overview of recent advancements and future perspectives in PE research specifically focused on plants, examining the key technical considerations of its applications.

show abstract

Section: Pe Applications In Plantsmentioning

confidence: 99%

Prime editing: Mechanism insight and recent applications in plants

Vu,

Nguyen,

Kim

et al. 2023

Plant Biotechnology Journal

View full text Add to dashboard Cite

show abstract

“…BEs and PEs can certainly contribute to the development of superior varieties with increased yields, improved nutrient content, broad adaptability in the environment, and increased efficiency in the use of agricultural inputs [ 199 , 200 , 201 ]. In 2022, Li et al achieved an editing efficiency of up to 66.7% using PE for precision editing at two adjacent target loci within the rice waxy gene [ 202 ]. In 2023, Qiao et al achieved simultaneous genetic precision editing of multiple genes and targets in maize with an optimized PE system [ 114 ].…”

Section: Discussion and Perspectivementioning

confidence: 99%

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Zhou

Luan

Liu

et al. 2023

Plants

View full text Add to dashboard Cite

Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature and is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation of transgene-free genome-edited plants in a short period and has the advantages of simplicity, high efficiency, high specificity, and low production costs, which greatly facilitate the study of gene functions. In plant molecular breeding, the gene-editing efficiency of the CRISPR-Cas9 system has proven to be a key step in influencing the effectiveness of molecular breeding, with improvements in gene-editing efficiency recently becoming a focus of reported scientific research. This review details strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding, including Cas9 variant enzyme engineering, the effect of multiple promoter driven Cas9, and gRNA efficient optimization and expression strategies. It also briefly introduces the optimization strategies of the CRISPR/Cas12a system and the application of BE and PE precision editing. These strategies are beneficial for the further development and optimization of gene editing systems in the field of plant molecular breeding.

show abstract

“…precision and flexibility in editing a gene than base editing and HDR (Anzalone et al, 2019). Successes have been reported in using PE for modifying genes in plant species, but the editing efficiency was quite low with the early versions of prime editors (Jiang et al, 2020;Li, Ma, et al, 2022;Lu et al, 2021;Perroud et al, 2022;Tang et al, 2020;Xu et al, 2020). Extensive efforts have been made to improve various components of PE for enhancing editing efficiency in both mammalian and plant systems (Chen and Liu, 2023).…”

Section: Introductionmentioning

confidence: 99%

“…The reverse transcriptase fused to nCas9 is able to repair the nicked target DNA sequence by using the RNA repair template of the pegRNA to precisely incorporate desired nucleotide changes to the genomic target and offers even greater precision and flexibility in editing a gene than base editing and HDR (Anzalone et al ., 2019). Successes have been reported in using PE for modifying genes in plant species, but the editing efficiency was quite low with the early versions of prime editors (Jiang et al ., 2020; Li, Li, et al ., 2020; Li, Ma, et al ., 2022; Li et al ., 2022; Lu et al ., 2021; Perroud et al ., 2022; Tang et al ., 2020; Xu et al ., 2020). Extensive efforts have been made to improve various components of PE for enhancing editing efficiency in both mammalian and plant systems (Chen and Liu, 2023).…”

Section: Introductionmentioning

confidence: 99%

Editing VvDXS1 for the creation of muscat flavour in Vitis vinifera cv. Scarlet Royal

Yang,

Wheatley,

Meakem

et al. 2024

Plant Biotechnology Journal

View full text Add to dashboard Cite

SummaryMuscat flavour represents a group of unique aromatic attributes in some grape varieties. Biochemically, grape berries with muscat flavour produce high levels of monoterpenes. Monoterpene biosynthesis is mainly through the DOXP/MEP pathway, and VvDXS1 encodes the first enzyme in this plastidial pathway of terpene biosynthesis in grapevine. A single‐point mutation resulting in the substitution of a lysine with an asparagine at position 284 in the VvDXS1 protein has previously been identified as the major cause for producing muscat flavour in grapes. In this study, the same substitution in the VvDXS1 protein was successfully created through prime editing in the table grape Vitis vinifera cv. ‘Scarlet Royal’. The targeted point mutation was detected in most of the transgenic vines, with varying editing efficiencies. No unintended mutations were detected in the edited alleles, either by PCR Sanger sequencing or by amplicon sequencing. More than a dozen edited vines were identified with an editing efficiency of more than 50%, indicating that these vines were likely derived from single cells in which one allele was edited. These vines had much higher levels of monoterpenes in their leaves than the control, similar to what was found in leaf samples between field‐grown muscat and non‐muscat grapes.

show abstract

A straightforward plant prime editing system enabled highly efficient precise editing of rice Waxy gene

Cited by 12 publications

References 36 publications

Prime editing: Mechanism insight and recent applications in plants

Prime editing: Mechanism insight and recent applications in plants

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Editing VvDXS1 for the creation of muscat flavour in Vitis vinifera cv. Scarlet Royal

Contact Info

Product

Resources

About