Correction to: Enhanced polyethylene glycol (PEG)–mediated protoplast transformation system for the phytopathogenic fungus, Ganoderma boninense

Abstract Background To date, CRISPR/Cas9 RNP editing tools have not been applied to the genetic modification of banana. Here, the establishment of a PEG-mediated banana protoplast transformation system makes it possible to build an efficient DNA-free method for a site-directed mutagenesis system. Results Protoplasts constitute a versatile platform for transient expression in plant science. In this study, we established a PEG-mediated banana protoplast transformation system. This system was further optimized for successfully delivering CRISPR/Cas9 and CRISPR/Cas12a plasmids and CRISPR/Cas9 ribonucleoproteins (RNPs) for targeted delivery of the PDS gene into banana protoplasts. Specific bands were observed in PCR-Restriction Enzyme Digestion (PCR-RE) assays, and Sanger sequencing of single clones further confirmed the occurrence of indels at target sites. Deep amplicon sequencing results showed that the editing efficiency of the CRISPR/Cas9 system was higher than that of the other two systems. Conclusions The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana. The application of the CRISPR/Cas9 RNP system enables the generation of banana plants engineered by DNA-free gene editing.

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An efficient polyethylene glycol-mediated transformation system of lentiviral vector in Shiraia bambusicola

Process Biochemistry ◽

10.1016/j.procbio.2016.07.013 ◽

2016 ◽

Vol 51 (10) ◽

pp. 1357-1362 ◽

Cited By ~ 10

Author(s):

Huaxiang Deng ◽

Ruijie Gao ◽

Jiajun Chen ◽

Xiangru Liao ◽

Yujie Cai

Keyword(s):

Polyethylene Glycol ◽

Lentiviral Vector ◽

Transformation System ◽

Shiraia Bambusicola

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Draft Genome Sequence of the Phytopathogenic Fungus Ganoderma boninense , the Causal Agent of Basal Stem Rot Disease on Oil Palm

Genome Announcements ◽

10.1128/genomea.00122-18 ◽

2018 ◽

Vol 6 (17) ◽

Cited By ~ 7

Author(s):

Condro Utomo ◽

Zulfikar Achmad Tanjung ◽

Redi Aditama ◽

Rika Fithri Nurani Buana ◽

Antonius Dony Madu Pratomo ◽

...

Keyword(s):

Fungal Pathogen ◽

Elaeis Guineensis ◽

Draft Genome ◽

Nuclear Genome ◽

Stem Rot ◽

Phytopathogenic Fungus ◽

Ganoderma Boninense ◽

Basal Stem Rot ◽

Content Type ◽

Rot Disease

ABSTRACT Ganoderma boninense is the dominant fungal pathogen of basal stem rot (BSR) disease on Elaeis guineensis . We sequenced the nuclear genome of mycelia using both Illumina and Pacific Biosciences platforms for assembly of scaffolds. The draft genome comprised 79.24 Mb, 495 scaffolds, and 26,226 predicted coding sequences.

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Establishment of A PEG-mediated Protoplast Transformation System Based on DNA and CRISPR/Cas9 Ribonucleoprotein Complexes For Banana

10.21203/rs.3.rs-32229/v1 ◽

2020 ◽

Author(s):

Shaoping Wu ◽

Haocheng Zhu ◽

Jinxing Liu ◽

Qiaosong Yang ◽

Xiuhong Shao ◽

...

Keyword(s):

Transient Expression ◽

Amplicon Sequencing ◽

Enzyme Digestion ◽

Site Directed Mutagenesis ◽

Plant Science ◽

Restriction Enzyme Digestion ◽

Protoplast Transformation ◽

Transformation System ◽

Ribonucleoprotein Complexes ◽

Target Sites

Abstract Background: To date, CRISPR/Cas9 RNPs editing tools have not been applied to genetic modification of banana. Here, the establishment of PEG-mediated banana protoplast transformation system makes it possible to build an efficient DNA- free method for the site-directed mutagenesis system.Results: Protoplasts are a versatile platform for transient expression in plant science. In this study, we established a PEG-mediated banana protoplast transformation system. This system was further optimized for successfully delivering plasmids of CRISPR/Cas9 and CRISPR/Cas12a , CRISPR/Cas9 ribonucleoproteins (RNPs) that targeted PDS gene into banana protoplasts. Specific bands were observed in PCR-Restriction Enzyme Digestion (PCR-RE) assays and monoclonal sequencing further confirmed the occurring of indels at target sites. Deep amplicon sequencing results showed that the editing efficiency of CRISPR/Cas9 system was higher than that of the other two systems. Conclusions: The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana.The application of CRISPR/Cas9 RNPs system enables the generation of DNA-free genome edited banana plants.

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