PoplarV1, Main, Exploration, bibRecord, 001D98

Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.

Identifieur interne : 001D98 ( Main/Exploration ); précédent : 001D97; suivant : 001D99

Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.

Auteurs : Di Fan [République populaire de Chine] ; Tingting Liu [République populaire de Chine] ; Chaofeng Li [République populaire de Chine] ; Bo Jiao [République populaire de Chine] ; Shuang Li [République populaire de Chine] ; Yishu Hou [République populaire de Chine] ; Keming Luo [République populaire de Chine]

Source :

Scientific reports [ 2045-2322 ] ; 2015.

RBID : pubmed:26193631

Descripteurs français

English descriptors

KwdEn :
- Alleles (MeSH), Base Sequence (MeSH), CRISPR-Cas Systems (genetics), Genes, Plant (MeSH), Heterozygote (MeSH), Homozygote (MeSH), Molecular Sequence Data (MeSH), Mutagenesis (genetics), Mutation (genetics), Phenotype (MeSH), Plants, Genetically Modified (MeSH), Populus (genetics), RNA Editing (genetics), RNA, Guide (genetics).
MESH :
- chemical , genetics : RNA, Guide.
- genetics : CRISPR-Cas Systems, Mutagenesis, Mutation, Populus, RNA Editing.
- Alleles, Base Sequence, Genes, Plant, Heterozygote, Homozygote, Molecular Sequence Data, Phenotype, Plants, Genetically Modified.

Abstract

Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been applied to edit the plant genome in several herbaceous plant species. However, it remains unknown whether this system can be used for genome editing in woody plants. In this study, we describe the genome editing and targeted gene mutation in a woody species, Populus tomentosa Carr. via the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed to target with distinct poplar genomic sites of the phytoene desaturase gene 8 (PtoPDS) which are followed by the protospacer-adjacent motif (PAM). After Agrobacterium-mediated transformation, obvious albino phenotype was observed in transgenic poplar plants. By analyzing the RNA-guided genome-editing events, 30 out of 59 PCR clones were homozygous mutants, 2 out of 59 were heterozygous mutants and the mutation efficiency at these target sites was estimated to be 51.7%. Our data demonstrate that the Cas9/sgRNA system can be exploited to precisely edit genomic sequence and effectively create knockout mutations in woody plants.

DOI: 10.1038/srep12217
PubMed: 26193631
PubMed Central: PMC4507398

Affiliations:

République populaire de Chine

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Le document en format XML

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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Alleles (MeSH)</term>
<term>Base Sequence (MeSH)</term>
<term>CRISPR-Cas Systems (genetics)</term>
<term>Genes, Plant (MeSH)</term>
<term>Heterozygote (MeSH)</term>
<term>Homozygote (MeSH)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Mutagenesis (genetics)</term>
<term>Mutation (genetics)</term>
<term>Phenotype (MeSH)</term>
<term>Plants, Genetically Modified (MeSH)</term>
<term>Populus (genetics)</term>
<term>RNA Editing (genetics)</term>
<term>RNA, Guide (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN guide (génétique)</term>
<term>Allèles (MeSH)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Gènes de plante (MeSH)</term>
<term>Homozygote (MeSH)</term>
<term>Hétérozygote (MeSH)</term>
<term>Mutagenèse (génétique)</term>
<term>Mutation (génétique)</term>
<term>Phénotype (MeSH)</term>
<term>Populus (génétique)</term>
<term>Systèmes CRISPR-Cas (génétique)</term>
<term>Séquence nucléotidique (MeSH)</term>
<term>Végétaux génétiquement modifiés (MeSH)</term>
<term>Édition des ARN (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Guide</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>CRISPR-Cas Systems</term>
<term>Mutagenesis</term>
<term>Mutation</term>
<term>Populus</term>
<term>RNA Editing</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN guide</term>
<term>Mutagenèse</term>
<term>Mutation</term>
<term>Populus</term>
<term>Systèmes CRISPR-Cas</term>
<term>Édition des ARN</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Alleles</term>
<term>Base Sequence</term>
<term>Genes, Plant</term>
<term>Heterozygote</term>
<term>Homozygote</term>
<term>Molecular Sequence Data</term>
<term>Phenotype</term>
<term>Plants, Genetically Modified</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Allèles</term>
<term>Données de séquences moléculaires</term>
<term>Gènes de plante</term>
<term>Homozygote</term>
<term>Hétérozygote</term>
<term>Phénotype</term>
<term>Séquence nucléotidique</term>
<term>Végétaux génétiquement modifiés</term>
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<front><div type="abstract" xml:lang="en">Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been applied to edit the plant genome in several herbaceous plant species. However, it remains unknown whether this system can be used for genome editing in woody plants. In this study, we describe the genome editing and targeted gene mutation in a woody species, Populus tomentosa Carr. via the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed to target with distinct poplar genomic sites of the phytoene desaturase gene 8 (PtoPDS) which are followed by the protospacer-adjacent motif (PAM). After Agrobacterium-mediated transformation, obvious albino phenotype was observed in transgenic poplar plants. By analyzing the RNA-guided genome-editing events, 30 out of 59 PCR clones were homozygous mutants, 2 out of 59 were heterozygous mutants and the mutation efficiency at these target sites was estimated to be 51.7%. Our data demonstrate that the Cas9/sgRNA system can be exploited to precisely edit genomic sequence and effectively create knockout mutations in woody plants. </div>
</front>
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<DateCompleted><Year>2016</Year>
<Month>08</Month>
<Day>02</Day>
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<DateRevised><Year>2018</Year>
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<Day>13</Day>
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<PubDate><Year>2015</Year>
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<ArticleTitle>Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.</ArticleTitle>
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<Abstract><AbstractText>Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been applied to edit the plant genome in several herbaceous plant species. However, it remains unknown whether this system can be used for genome editing in woody plants. In this study, we describe the genome editing and targeted gene mutation in a woody species, Populus tomentosa Carr. via the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed to target with distinct poplar genomic sites of the phytoene desaturase gene 8 (PtoPDS) which are followed by the protospacer-adjacent motif (PAM). After Agrobacterium-mediated transformation, obvious albino phenotype was observed in transgenic poplar plants. By analyzing the RNA-guided genome-editing events, 30 out of 59 PCR clones were homozygous mutants, 2 out of 59 were heterozygous mutants and the mutation efficiency at these target sites was estimated to be 51.7%. Our data demonstrate that the Cas9/sgRNA system can be exploited to precisely edit genomic sequence and effectively create knockout mutations in woody plants. </AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Fan</LastName>
<ForeName>Di</ForeName>
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</AffiliationInfo>
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<ForeName>Tingting</ForeName>
<Initials>T</Initials>
<AffiliationInfo><Affiliation>Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Transgenic Plant and Safety Control, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing 400715, China.</Affiliation>
</AffiliationInfo>
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<ForeName>Chaofeng</ForeName>
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<Author ValidYN="Y"><LastName>Jiao</LastName>
<ForeName>Bo</ForeName>
<Initials>B</Initials>
<AffiliationInfo><Affiliation>Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Transgenic Plant and Safety Control, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing 400715, China.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Li</LastName>
<ForeName>Shuang</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Transgenic Plant and Safety Control, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing 400715, China.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Hou</LastName>
<ForeName>Yishu</ForeName>
<Initials>Y</Initials>
<AffiliationInfo><Affiliation>Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Transgenic Plant and Safety Control, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing 400715, China.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Luo</LastName>
<ForeName>Keming</ForeName>
<Initials>K</Initials>
<AffiliationInfo><Affiliation>1] Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Transgenic Plant and Safety Control, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing 400715, China [2] Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, 810008 Xining, China.</Affiliation>
</AffiliationInfo>
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<Language>eng</Language>
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<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
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<Month>07</Month>
<Day>20</Day>
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Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.

Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.

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