Evaluating the Efficiency of gRNAs in CRISPR/Cas9 Mediated Genome Editing in Poplars.
Identifieur interne : 000981 ( Main/Exploration ); précédent : 000980; suivant : 000982Evaluating the Efficiency of gRNAs in CRISPR/Cas9 Mediated Genome Editing in Poplars.
Auteurs : Tobias Bruegmann [Allemagne] ; Khira Deecke [Allemagne] ; Matthias Fladung [Allemagne]Source :
- International journal of molecular sciences [ 1422-0067 ] ; 2019.
Descripteurs français
- KwdFr :
- MESH :
- génétique : ARN guide, Génome végétal, Populus, Systèmes CRISPR-Cas.
- Agrobacterium, Édition de gène.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : RNA, Guide.
- genetics : CRISPR-Cas Systems, Genome, Plant, Populus.
- Agrobacterium, Gene Editing.
Abstract
CRISPR/Cas9 has become one of the most promising techniques for genome editing in plants and works very well in poplars with an Agrobacterium-mediated transformation system. We selected twelve genes, including SOC1, FUL, and their paralogous genes, four NFP-like genes and TOZ19 for three different research topics. The gRNAs were designed for editing, and, together with a constitutively expressed Cas9 nuclease, transferred either into the poplar hybrid Populus × canescens or into P. tremula. The regenerated lines showed different types of editing and revealed several homozygous editing events which are of special interest in perennial species because of limited back-cross ability. Through a time series, we could show that despite the constitutive expression of the Cas9 nuclease, no secondary editing of the target region occurred. Thus, constitutive Cas9 expression does not seem to pose any risk to additional editing events. Based on various criteria, we obtained evidence for a relationship between the structure of gRNA and the efficiency of gene editing. In particular, the GC content, purine residues in the gRNA end, and the free accessibility of the seed region seemed to be highly important for genome editing in poplars. Based on our findings on nine different poplar genes, efficient gRNAs can be designed for future efficient editing applications in poplars.
DOI: 10.3390/ijms20153623
PubMed: 31344908
PubMed Central: PMC6696231
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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type="eISSN">1422-0067</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium (MeSH)</term><term>CRISPR-Cas Systems (genetics)</term><term>Gene Editing (MeSH)</term><term>Genome, Plant (genetics)</term><term>Populus (genetics)</term><term>RNA, Guide (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN guide (génétique)</term><term>Agrobacterium (MeSH)</term><term>Génome végétal (génétique)</term><term>Populus (génétique)</term><term>Systèmes CRISPR-Cas (génétique)</term><term>Édition de gène (MeSH)</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>Genome, Plant</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN guide</term><term>Génome végétal</term><term>Populus</term><term>Systèmes CRISPR-Cas</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Agrobacterium</term><term>Gene Editing</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Agrobacterium</term><term>Édition de gène</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">CRISPR/Cas9 has become one of the most promising techniques for genome editing in plants and works very well in poplars with an <i>Agrobacterium</i>-mediated transformation system. We selected twelve genes, including <i>SOC1</i>, <i>FUL</i>, and their paralogous genes, four <i>NFP-like</i> genes and <i>TOZ19</i> for three different research topics. The gRNAs were designed for editing, and, together with a constitutively expressed Cas9 nuclease, transferred either into the poplar hybrid <i>Populus</i> × <i>canescens</i> or into <i>P. tremula</i>. The regenerated lines showed different types of editing and revealed several homozygous editing events which are of special interest in perennial species because of limited back-cross ability. Through a time series, we could show that despite the constitutive expression of the Cas9 nuclease, no secondary editing of the target region occurred. Thus, constitutive Cas9 expression does not seem to pose any risk to additional editing events. Based on various criteria, we obtained evidence for a relationship between the structure of gRNA and the efficiency of gene editing. In particular, the GC content, purine residues in the gRNA end, and the free accessibility of the seed region seemed to be highly important for genome editing in poplars. Based on our findings on nine different poplar genes, efficient gRNAs can be designed for future efficient editing applications in poplars.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">31344908</PMID><DateCompleted><Year>2019</Year><Month>12</Month><Day>27</Day></DateCompleted><DateRevised><Year>2020</Year><Month>02</Month><Day>25</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1422-0067</ISSN><JournalIssue CitedMedium="Internet"><Volume>20</Volume><Issue>15</Issue><PubDate><Year>2019</Year><Month>Jul</Month><Day>24</Day></PubDate></JournalIssue><Title>International journal of molecular sciences</Title><ISOAbbreviation>Int J Mol Sci</ISOAbbreviation></Journal><ArticleTitle>Evaluating the Efficiency of gRNAs in CRISPR/Cas9 Mediated Genome Editing in Poplars.</ArticleTitle><ELocationID EIdType="pii" ValidYN="Y">E3623</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.3390/ijms20153623</ELocationID><Abstract><AbstractText>CRISPR/Cas9 has become one of the most promising techniques for genome editing in plants and works very well in poplars with an <i>Agrobacterium</i>-mediated transformation system. We selected twelve genes, including <i>SOC1</i>, <i>FUL</i>, and their paralogous genes, four <i>NFP-like</i> genes and <i>TOZ19</i> for three different research topics. The gRNAs were designed for editing, and, together with a constitutively expressed Cas9 nuclease, transferred either into the poplar hybrid <i>Populus</i> × <i>canescens</i> or into <i>P. tremula</i>. The regenerated lines showed different types of editing and revealed several homozygous editing events which are of special interest in perennial species because of limited back-cross ability. Through a time series, we could show that despite the constitutive expression of the Cas9 nuclease, no secondary editing of the target region occurred. Thus, constitutive Cas9 expression does not seem to pose any risk to additional editing events. Based on various criteria, we obtained evidence for a relationship between the structure of gRNA and the efficiency of gene editing. In particular, the GC content, purine residues in the gRNA end, and the free accessibility of the seed region seemed to be highly important for genome editing in poplars. Based on our findings on nine different poplar genes, efficient gRNAs can be designed for future efficient editing applications in poplars.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bruegmann</LastName><ForeName>Tobias</ForeName><Initials>T</Initials><Identifier Source="ORCID">0000-0001-9823-5630</Identifier><AffiliationInfo><Affiliation>Thuenen Institute of Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany. tobias.bruegmann@thuenen.de.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Deecke</LastName><ForeName>Khira</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Thuenen Institute of Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fladung</LastName><ForeName>Matthias</ForeName><Initials>M</Initials><Identifier Source="ORCID">0000-0001-9301-8581</Identifier><AffiliationInfo><Affiliation>Thuenen Institute of Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany. matthias.fladung@thuenen.de.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>0315972A, 031B0203B, 031B0535</GrantID><Agency>Bundesministerium für Bildung und Forschung</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>07</Month><Day>24</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Int J Mol Sci</MedlineTA><NlmUniqueID>101092791</NlmUniqueID><ISSNLinking>1422-0067</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017394">RNA, Guide</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D060054" 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