Prime Editing in the model plant Physcomitrium patens and its potential in the tetraploid potato

Perroud, Pierre‐François; Guyon‐Debast, Anouchka; Veillet, Florian; Kermarrec, Marie-Paule; Chauvin, Laura; Chauvin, Jean-Éric; Gallois, Jean‐Luc; Nogué, Fabien

doi:10.1016/j.plantsci.2021.111162

Cited by 42 publications

(43 citation statements)

References 55 publications

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“…In addition, PEs have been found to successfully introduce insertions, deletions, transitions, and transversions (Anzalone et al, 2019). Perroud et al (2022) reported that 0.06% of transformed protoplasts of Physcomitrium were edited, which is less than the standard Cas9 mediated and base editing mutagenic strategies. However, the edit's specificity is higher than CRISPR/Cas systems, and off-targets are few or none.…”

Section: Prime Editingmentioning

confidence: 99%

“…Agrobacterium -mediated transformation and floral dip agroinfiltration are the preferred modes of gene transfer as single copy inserts are efficiently achieved. However, other methods such as electroporation, PEG-mediated gene uptake, microinjection, and particle bombardment have been tested in different plants and are now expanding rapidly to include monocots (rice and maize), dicots ( Arabidopsis , Nicotiana benthamiana , potato, and tomato), and even the bryophyte, Physcomitrium patens ( Perroud et al, 2022 ) that is well known for incorporating DNA into specific genomic sites due to its innately high frequencies of homologous recombination ( Rensing et al, 2020 ).…”

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

“…The targeted sites are usually PCR amplified from the genomic DNA isolated from transformed plants and sequenced to identify the edits. Most researchers have done Sanger’s sequencing, although the HRM-High Resolution Melting analysis has been included before sequencing by Perroud et al (2022) . Hi-TOM (high-throughput tracking of mutations) was used by Xu et al (2022) in maize and rice.…”

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

“…Different selection and counter-selection strategies have been tested for the selection of transformed/edited cells. Perroud et al (2022) have tested the use of APT/APRT (adenine phosphoribosyl transferase) enzyme that catalyzes the conversion of adenine to AMP in Physcomitrium . This enzyme can convert 2-fluoroadenine (2FA) supplemented in the culture medium into a toxic 2-fluoro AMP counter selective compound.…”

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

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Comprehending the evolution of gene editing platforms for crop trait improvement

et al. 2022

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CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas (CRISPR-associated) system was initially discovered as an underlying mechanism for conferring adaptive immunity to bacteria and archaea against viruses. Over the past decade, this has been repurposed as a genome-editing tool. Numerous gene editing-based crop improvement technologies involving CRISPR/Cas platforms individually or in combination with next-generation sequencing methods have been developed that have revolutionized plant genome-editing methodologies. Initially, CRISPR/Cas nucleases replaced the earlier used sequence-specific nucleases (SSNs), such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), to address the problem of associated off-targets. The adaptation of this platform led to the development of concepts such as epigenome editing, base editing, and prime editing. Epigenome editing employed epi-effectors to manipulate chromatin structure, while base editing uses base editors to engineer precise changes for trait improvement. Newer technologies such as prime editing have now been developed as a “search-and-replace” tool to engineer all possible single-base changes. Owing to the availability of these, the field of genome editing has evolved rapidly to develop crop plants with improved traits. In this review, we present the evolution of the CRISPR/Cas system into new-age methods of genome engineering across various plant species and the impact they have had on tweaking plant genomes and associated outcomes on crop improvement initiatives.

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Section: Prime Editingmentioning

confidence: 99%

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

Section: Evolution Of Clustered Regularly Interspaced Short Palindrom...mentioning

confidence: 99%

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Comprehending the evolution of gene editing platforms for crop trait improvement

et al. 2022

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“…Prime editing was first described at the end of 2019 in human and yeast cells ( Anzalone et al, 2019 ) and since then it has been used broadly in several plant species ( Butt et al, 2020 ; Hua et al, 2020 ; Jiang et al, 2020 ; Li et al, 2020b ; Lin et al, 2020 ; Xu et al, 2020a , b ; Lu et al, 2021 ; Perroud et al, 2022 ). PE is based on a “search and replace” approach in which a nickase Cas9 (nCas9) is fused to a reverse transcriptase (RT), coupled to a prime editing guide RNA (pegRNA) that specifies both the target site and provides the template for the edition ( Figure 2 ).…”

Section: Prime Editingmentioning

confidence: 99%

Molecular and Computational Strategies to Increase the Efficiency of CRISPR-Based Techniques

et al. 2022

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The prokaryote-derived Clustered Regularly Interspaced Palindromic Repeats (CRISPR)/Cas mediated gene editing tools have revolutionized our ability to precisely manipulate specific genome sequences in plants and animals. The simplicity, precision, affordability, and robustness of this technology have allowed a myriad of genomes from a diverse group of plant species to be successfully edited. Even though CRISPR/Cas, base editing, and prime editing technologies have been rapidly adopted and implemented in plants, their editing efficiency rate and specificity varies greatly. In this review, we provide a critical overview of the recent advances in CRISPR/Cas9-derived technologies and their implications on enhancing editing efficiency. We highlight the major efforts of engineering Cas9, Cas12a, Cas12b, and Cas12f proteins aiming to improve their efficiencies. We also provide a perspective on the global future of agriculturally based products using DNA-free CRISPR/Cas techniques. The improvement of CRISPR-based technologies efficiency will enable the implementation of genome editing tools in a variety of crop plants, as well as accelerate progress in basic research and molecular breeding.

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Base Editing and Prime Editing

Kaya

2023

A Roadmap for Plant Genome Editing

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The development of new adaptations of CRISPR-based genome editing platforms, such as base editing and prime editing, made it possible to broaden the scope and applications of genome editing in plants. First base editing and, more recently, prime editing evade the creation of double-stranded breaks in deoxyribonucleic acid (DNA) and the requirement of donor template of DNA for repair while enhancing editing efficiency and product purity over CRISPR/Cas9. As base-pair changes in genomic DNA determine many significant agronomic traits, crop varieties can be developed by precisely converting specific single bases in plant genomes. While base editing can introduce specific nucleotide changes, such as transition and transversion mutations in the targeted region, prime editing can create precise insertions, deletions, and all 12 types of point mutations using the “search-and-replace” method.This chapter provides the basic principles of base editing and prime editing technologies and their practical applications in plants. The chapter also summarizes the recent breakthroughs in applying base and prime editors in diverse plant species, including their use in improving disease resistance, herbicide resistance, nutritional quality, crop yield, and quality. Finally, this chapter aims to clearly understand base editing and prime editing in plants by outlining potential developments.

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Prime Editing in the model plant Physcomitrium patens and its potential in the tetraploid potato

Cited by 42 publications

References 55 publications

Comprehending the evolution of gene editing platforms for crop trait improvement

Comprehending the evolution of gene editing platforms for crop trait improvement

Molecular and Computational Strategies to Increase the Efficiency of CRISPR-Based Techniques

Base Editing and Prime Editing

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