CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.
Identifieur interne : 000001 ( Main/Exploration ); précédent : 000000; suivant : 000002CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.
Auteurs : Veerendra Sharma [États-Unis] ; Wenguang Zheng [États-Unis] ; Jun Huang [États-Unis] ; David E. Cook [États-Unis]Source :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2021.
Abstract
Application of the CRISPR-Cas prokaryotic immune system for single-stranded RNA targeting will have significant impacts on RNA analysis and engineering. The class 2 Type VI CRISPR-Cas13 system is an RNA-guided RNA-nuclease system capable of binding and cleaving target single-stranded RNA substrates in a sequence-specific manner. In addition to RNA interference, the Cas13a system has application from manipulating RNA modifications, to editing RNA sequence, to use as a nucleic acid detection tool. This protocol uses the Cas13a ortholog from Leptotrichia buccalis for transient expression in plant cells providing antiviral defense. We cover all the necessary information for cloning the Cas13 protein, crRNA guide cassette, performing transient Agrobacterium-mediated expression of the necessary Cas13a components and target RNA-virus, visualization of virus infection, and molecular quantification of viral accumulation using quantitative PCR.
DOI: 10.1007/978-1-0716-0743-5_1
PubMed: 32797447
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.</title><author><name sortKey="Sharma, Veerendra" sort="Sharma, Veerendra" uniqKey="Sharma V" first="Veerendra" last="Sharma">Veerendra Sharma</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Zheng, Wenguang" sort="Zheng, Wenguang" uniqKey="Zheng W" first="Wenguang" last="Zheng">Wenguang Zheng</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Huang, Jun" sort="Huang, Jun" uniqKey="Huang J" first="Jun" last="Huang">Jun Huang</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Cook, David E" sort="Cook, David E" uniqKey="Cook D" first="David E" last="Cook">David E. Cook</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA. decook@ksu.edu.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2021">2021</date><idno type="RBID">pubmed:32797447</idno><idno type="pmid">32797447</idno><idno type="doi">10.1007/978-1-0716-0743-5_1</idno><idno type="wicri:Area/Main/Corpus">000020</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000020</idno><idno type="wicri:Area/Main/Curation">000020</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000020</idno><idno type="wicri:Area/Main/Exploration">000020</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.</title><author><name sortKey="Sharma, Veerendra" sort="Sharma, Veerendra" uniqKey="Sharma V" first="Veerendra" last="Sharma">Veerendra Sharma</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Zheng, Wenguang" sort="Zheng, Wenguang" uniqKey="Zheng W" first="Wenguang" last="Zheng">Wenguang Zheng</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Huang, Jun" sort="Huang, Jun" uniqKey="Huang J" first="Jun" last="Huang">Jun Huang</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author><author><name sortKey="Cook, David E" sort="Cook, David E" uniqKey="Cook D" first="David E" last="Cook">David E. Cook</name><affiliation wicri:level="2"><nlm:affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA. decook@ksu.edu.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Kansas State University, Manhattan, KS</wicri:regionArea><placeName><region type="state">Kansas</region></placeName></affiliation></author></analytic><series><title level="j">Methods in molecular biology (Clifton, N.J.)</title><idno type="eISSN">1940-6029</idno><imprint><date when="2021" type="published">2021</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Application of the CRISPR-Cas prokaryotic immune system for single-stranded RNA targeting will have significant impacts on RNA analysis and engineering. The class 2 Type VI CRISPR-Cas13 system is an RNA-guided RNA-nuclease system capable of binding and cleaving target single-stranded RNA substrates in a sequence-specific manner. In addition to RNA interference, the Cas13a system has application from manipulating RNA modifications, to editing RNA sequence, to use as a nucleic acid detection tool. This protocol uses the Cas13a ortholog from Leptotrichia buccalis for transient expression in plant cells providing antiviral defense. We cover all the necessary information for cloning the Cas13 protein, crRNA guide cassette, performing transient Agrobacterium-mediated expression of the necessary Cas13a components and target RNA-virus, visualization of virus infection, and molecular quantification of viral accumulation using quantitative PCR.</div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">32797447</PMID><DateRevised><Year>2020</Year><Month>08</Month><Day>17</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1940-6029</ISSN><JournalIssue CitedMedium="Internet"><Volume>2170</Volume><PubDate><Year>2021</Year></PubDate></JournalIssue><Title>Methods in molecular biology (Clifton, N.J.)</Title><ISOAbbreviation>Methods Mol Biol</ISOAbbreviation></Journal><ArticleTitle>CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.</ArticleTitle><Pagination><MedlinePgn>1-18</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/978-1-0716-0743-5_1</ELocationID><Abstract><AbstractText>Application of the CRISPR-Cas prokaryotic immune system for single-stranded RNA targeting will have significant impacts on RNA analysis and engineering. The class 2 Type VI CRISPR-Cas13 system is an RNA-guided RNA-nuclease system capable of binding and cleaving target single-stranded RNA substrates in a sequence-specific manner. In addition to RNA interference, the Cas13a system has application from manipulating RNA modifications, to editing RNA sequence, to use as a nucleic acid detection tool. This protocol uses the Cas13a ortholog from Leptotrichia buccalis for transient expression in plant cells providing antiviral defense. We cover all the necessary information for cloning the Cas13 protein, crRNA guide cassette, performing transient Agrobacterium-mediated expression of the necessary Cas13a components and target RNA-virus, visualization of virus infection, and molecular quantification of viral accumulation using quantitative PCR.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Sharma</LastName><ForeName>Veerendra</ForeName><Initials>V</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zheng</LastName><ForeName>Wenguang</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Jun</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Cook</LastName><ForeName>David E</ForeName><Initials>DE</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Kansas State University, Manhattan, KS, USA. decook@ksu.edu.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, Non-P.H.S.</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Methods Mol Biol</MedlineTA><NlmUniqueID>9214969</NlmUniqueID><ISSNLinking>1064-3745</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Antiviral protection</Keyword><Keyword MajorTopicYN="Y">CRISPR-Cas13</Keyword><Keyword MajorTopicYN="Y">Plant biotechnology</Keyword><Keyword MajorTopicYN="Y">Plant virus</Keyword><Keyword MajorTopicYN="Y">RNA editing</Keyword><Keyword MajorTopicYN="Y">RNA targeting</Keyword><Keyword MajorTopicYN="Y">Transcriptome editing</Keyword><Keyword MajorTopicYN="Y">mRNA interference</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>8</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>8</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>8</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32797447</ArticleId><ArticleId IdType="doi">10.1007/978-1-0716-0743-5_1</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country><region><li>Kansas</li></region></list><tree><country name="États-Unis"><region name="Kansas"><name sortKey="Sharma, Veerendra" sort="Sharma, Veerendra" uniqKey="Sharma V" first="Veerendra" last="Sharma">Veerendra Sharma</name></region><name sortKey="Cook, David E" sort="Cook, David E" uniqKey="Cook D" first="David E" last="Cook">David E. Cook</name><name sortKey="Huang, Jun" sort="Huang, Jun" uniqKey="Huang J" first="Jun" last="Huang">Jun Huang</name><name sortKey="Zheng, Wenguang" sort="Zheng, Wenguang" uniqKey="Zheng W" first="Wenguang" last="Zheng">Wenguang Zheng</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Bois/explor/AgrobacTransV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000001 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000001 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Bois
|area= AgrobacTransV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:32797447
|texte= CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:32797447" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a AgrobacTransV1
| This area was generated with Dilib version V0.6.38. |  |