CRISPR/Cpf1-mediated DNA-free plant genome editing

Kim, Hye‐Ran; Kim, Sang‐Tae; Ryu, Jae‐Chun; Kang, Beum‐Chang; Kim, Jin-Soo

doi:10.1038/ncomms14406

Cited by 421 publications

(315 citation statements)

References 23 publications

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“…Cpf1 from Acidaminococcus sp. (AsCpf1) and Lachnospiraceae bacterium (LbCpf1) have been shown to be functional in rice, soybean and tobacco [34,[56][57][58]. In these plants, LbCpf1 mutagenesis efficiency, which seems to be correlated with the target sequence [56], appeared always higher than those of AsCpf1, reaching sometimes 100% in rice [58].…”

Section: Cas-like Proteinsmentioning

confidence: 99%

“…This strategy could soon be applied to many other crops amenable to biolistic delivery such as wheat, barley, sorghum, rice and soybean. RNP complexes formed between Cpf1 and adapted crRNAs were also tested in soybean targeting two endogenous genes [57]. Mutagenesis frequencies ranged from 0.0 to 11.7% using LbCpf1, and from 0.0 to 1.6% using AsCpf1.…”

Section: Transient Transformationmentioning

confidence: 99%

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Towards mastering CRISPR-induced gene knock-in in plants: Survey of key features and focus on the model Physcomitrella patens

Collonnier

Guyon‐Debast

Maclot

et al. 2017

Methods

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a b s t r a c tBeyond its predominant role in human and animal therapy, the CRISPR-Cas9 system has also become an essential tool for plant research and plant breeding. Agronomic applications rely on the mastery of gene inactivation and gene modification. However, if the knock-out of genes by non-homologous end-joining (NHEJ)-mediated repair of the targeted double-strand breaks (DSBs) induced by the CRISPR-Cas9 system is rather well mastered, the knock-in of genes by homology-driven repair or end-joining remains difficult to perform efficiently in higher plants. In this review, we describe the different approaches that can be tested to improve the efficiency of CRISPR-induced gene modification in plants, which include the use of optimal transformation and regeneration protocols, the design of appropriate guide RNAs and donor templates and the choice of nucleases and means of delivery. We also present what can be done to orient DNA repair pathways in the target cells, and we show how the moss Physcomitrella patens can be used as a model plant to better understand what DNA repair mechanisms are involved, and how this knowledge could eventually be used to define more performant strategies of CRISPR-induced gene knock-in.

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Section: Cas-like Proteinsmentioning

confidence: 99%

Section: Transient Transformationmentioning

confidence: 99%

Towards mastering CRISPR-induced gene knock-in in plants: Survey of key features and focus on the model Physcomitrella patens

Collonnier

Guyon‐Debast

Maclot

et al. 2017

Methods

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“…In a study, Song et al successfully corrected b-thalassemia mutations in iPSCs using CRISPR/Cas9 [27]. The recent success has been made when a DMD mouse model edited by CRISPR/Cas9 restored expression of the dystrophin protein and improved muscle pathology and strength [28].editing tools to emerge, including non-Cas9 based type II systems such as the recently described RNA-guided endonuclease Cpf1 and peptide nucleic acid (PNA) molecules [29,30]. Recently, Cpf1, a type V CRISPR effector has been successfully used to induce mutation in soybean genome [29].…”

Section: Crispr/cas9 and Gene Editingmentioning

confidence: 99%

Application of CRISPR/Cas9 Genome Editing in Genetic Disorders: A Systematic Review Up to Date

Pandey¹,

Tripathi²,

Bhushan³

et al. 2017

J Genet Syndr Gene Ther

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Genetic diseases in human are associated with congenital disorders and phenotypic traits. A single mutation in a gene can cause physical or mental problems, and sometimes both. Some diseases can be lethal, and there are still no cures for many of them. Socioeconomic burden of rare genetic diseases are increasing worldwide that have been tried to cure using various methods. However, they were not very successful till now. Genome editing technologies over the past few years is providing fast and effective tool to precisely manipulate the genome at specific locations. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated Cas9 (CRISPR/Cas9) system has been using from last few years in the field of biomedical research. CRISPR/Cas9 has advantages in terms of clinical applicability to treat genetic diseases like DMD, Hemophilia, β-Thalassemia and cystic fibrosis etc. and even in some cases this tool has already been successfully applied. Nevertheless, developed technologies for addition or deletion of genes have made notable progress in last few years and demonstrate some promising clinical results. However, several challenges still remain. Here, the latest applications of CRISPR-Cas9 technology in genetic disorders, current challenges and future directions are reviewed and discussed.

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“…RNP-mediated genome engineering was first shown in mammalian cells [56,57] but has since been demonstrated in many plant species [58][59][60][61]. Purified Cas9 protein is commercially available or can be overexpressed in Escherichia coli ( Figure 5).…”

Section: Tools For Dna-free Engineeringmentioning

confidence: 99%

“…Mutations at the target were found in 46% of callus tissues regenerated from lettuce protoplasts. Kim et al [61] used LbCpf1 and AsCpf1 RNPs to induce targeted mutagenesis in protoplasts of soybean and tobacco ( Figure 5B). Svitashev et al [59] and Liang et al [60] delivered RNPs into embryo cells of maize and wheat, respectively, using particle bombardment.…”

Section: Tools For Dna-free Engineeringmentioning

confidence: 99%

CRISPR-based tools for plant genome engineering

Silva

Patron

2017

Emerging Topics in Life Sciences

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Molecular tools adapted from bacterial CRISPR (clustered regulatory interspaced short palindromic repeat) adaptive immune systems have been demonstrated in an increasingly wide range of plant species. They have been applied for the induction of targeted mutations in one or more genes as well as for directing the integration of new DNA to specific genomic loci. The construction of molecular tools for multiplexed CRISPR-mediated editing in plants has been facilitated by cloning techniques that allow multiple sequences to be assembled together in a single cloning reaction. Modifications of the canonical Cas9 protein from Streptococcus pyogenes and the use of nucleases from other bacteria have increased the diversity of genomic sequences that can be targeted and allow the delivery of protein cargos such as transcriptional activators and repressors. Furthermore, the direct delivery of protein-RNA complexes to plant cells and tissues has enabled the production of engineered plants without the delivery or genomic integration of foreign DNA. Here, we review toolkits derived from bacterial CRISPR systems for targeted mutagenesis, gene delivery and modulation of gene expression in plants, focusing on their composition and the strategies employed to reprogramme them for the recognition of specific genomic targets.

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CRISPR/Cpf1-mediated DNA-free plant genome editing

Cited by 421 publications

References 23 publications

Towards mastering CRISPR-induced gene knock-in in plants: Survey of key features and focus on the model Physcomitrella patens

Towards mastering CRISPR-induced gene knock-in in plants: Survey of key features and focus on the model Physcomitrella patens

Application of CRISPR/Cas9 Genome Editing in Genetic Disorders: A Systematic Review Up to Date

CRISPR-based tools for plant genome engineering

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