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Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9

Bananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In thi... Full description

Journal Title: Transgenic Research 2018, Vol.27 (5), p.451-460
Main Author: Naim, Fatima
Other Authors: Dugdale, Benjamin , Kleidon, Jennifer , Brinin, Anthony , Shand, Kylie , Waterhouse, Peter , Dale, James
Format: Electronic Article Electronic Article
Language: English
Subjects:
PDS
RNA
Publisher: Cham: Springer International Publishing
ID: ISSN: 0962-8819
Link: https://www.ncbi.nlm.nih.gov/pubmed/29987710
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title: Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
format: Article
creator:
  • Naim, Fatima
  • Dugdale, Benjamin
  • Kleidon, Jennifer
  • Brinin, Anthony
  • Shand, Kylie
  • Waterhouse, Peter
  • Dale, James
subjects:
  • Albinism
  • Alleles
  • Allelomorphism
  • Animal Genetics and Genomics
  • beverages
  • Biomedical and Life Sciences
  • Biomedical Engineering/Biotechnology
  • Brief Communication
  • Cas9
  • Cavendish banana
  • CRISPR
  • CRISPR-Cas Systems
  • Desaturase
  • Disease resistance
  • food
  • Food sources
  • Gene Editing - methods
  • Genetic Engineering
  • Genetically modified organisms
  • Genome editing
  • Genomes
  • Genomics
  • Genotyping
  • gRNA
  • Life Sciences
  • Molecular Medicine
  • Musa - genetics
  • Oxidoreductases - genetics
  • PDS
  • Phenotypes
  • Plant Genetics and Genomics
  • Plant Proteins - genetics
  • Plants, Genetically Modified
  • Ribonucleic acid
  • RNA
  • RNA, Guide
  • Transgenics
ispartof: Transgenic Research, 2018, Vol.27 (5), p.451-460
description: Bananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase ( PDS ) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future.
language: eng
source:
identifier: ISSN: 0962-8819
fulltext: no_fulltext
issn:
  • 0962-8819
  • 1573-9368
url: Link


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titleGene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
creatorNaim, Fatima ; Dugdale, Benjamin ; Kleidon, Jennifer ; Brinin, Anthony ; Shand, Kylie ; Waterhouse, Peter ; Dale, James
creatorcontribNaim, Fatima ; Dugdale, Benjamin ; Kleidon, Jennifer ; Brinin, Anthony ; Shand, Kylie ; Waterhouse, Peter ; Dale, James
descriptionBananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase ( PDS ) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future.
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subjectAlbinism ; Alleles ; Allelomorphism ; Animal Genetics and Genomics ; beverages ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Brief Communication ; Cas9 ; Cavendish banana ; CRISPR ; CRISPR-Cas Systems ; Desaturase ; Disease resistance ; food ; Food sources ; Gene Editing - methods ; Genetic Engineering ; Genetically modified organisms ; Genome editing ; Genomes ; Genomics ; Genotyping ; gRNA ; Life Sciences ; Molecular Medicine ; Musa - genetics ; Oxidoreductases - genetics ; PDS ; Phenotypes ; Plant Genetics and Genomics ; Plant Proteins - genetics ; Plants, Genetically Modified ; Ribonucleic acid ; RNA ; RNA, Guide ; Transgenics
ispartofTransgenic Research, 2018, Vol.27 (5), p.451-460
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descriptionBananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase ( PDS ) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future.
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27Oxidoreductases - genetics
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29Phenotypes
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35RNA, Guide
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titleGene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
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29Phenotypes
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abstractBananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase ( PDS ) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future.
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