Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing
The CRISPR‐Cas9 gene editing system requires Cas9 endonuclease and guide RNAs (either the natural dual RNA consisting of crRNA and tracrRNA or a chimeric single guide RNA) that direct site‐specific double‐stranded DNA cleavage. This communication describes a click ligation approach that uses alkyne–...
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| Veröffentlicht in: | Chembiochem : a European journal of chemical biology 2016-10, Vol.17 (19), p.1809-1812 |
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| container_title | Chembiochem : a European journal of chemical biology |
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| creator | He, Kaizhang Chou, Eldon T. Begay, Shawn Anderson, Emily M. van Brabant Smith, Anja |
| description | The CRISPR‐Cas9 gene editing system requires Cas9 endonuclease and guide RNAs (either the natural dual RNA consisting of crRNA and tracrRNA or a chimeric single guide RNA) that direct site‐specific double‐stranded DNA cleavage. This communication describes a click ligation approach that uses alkyne–azide cycloaddition to generate a triazole‐linked single guide RNA (sgRNA). The conjugated sgRNA shows efficient and comparable genome editing activity to natural dual RNA and unmodified sgRNA constructs.
Editing efficiency: A triazole‐linked single guide RNA (sgRNA) was constructed by using click chemistry. The sgRNA showed efficient CRISPR‐Cas9 genome editing activity comparable to that of natural dual RNA and unmodified sgRNA constructs. These findings open new avenues for generating long sgRNA libraries in array or plate formats. |
| doi_str_mv | 10.1002/cbic.201600320 |
| format | Article |
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Editing efficiency: A triazole‐linked single guide RNA (sgRNA) was constructed by using click chemistry. The sgRNA showed efficient CRISPR‐Cas9 genome editing activity comparable to that of natural dual RNA and unmodified sgRNA constructs. These findings open new avenues for generating long sgRNA libraries in array or plate formats.</description><identifier>ISSN: 1439-4227</identifier><identifier>EISSN: 1439-7633</identifier><identifier>DOI: 10.1002/cbic.201600320</identifier><identifier>PMID: 27441384</identifier><language>eng</language><publisher>Germany: Blackwell Publishing Ltd</publisher><subject>Communication ; Communications ; conjugation ; CRISPR-Cas Systems - genetics ; CRISPR-Cas9 ; Editing ; Gene Editing ; gene technology ; Molecular Structure ; oligonucleotides ; RNA, Guide, CRISPR-Cas Systems - chemistry ; RNA, Guide, CRISPR-Cas Systems - metabolism ; single guide RNA ; Triazoles - chemistry ; Triazoles - metabolism</subject><ispartof>Chembiochem : a European journal of chemical biology, 2016-10, Vol.17 (19), p.1809-1812</ispartof><rights>2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.</rights><rights>2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5760-7de6d90f4a9ed1eced487c03671bce98eb1128a2682b30f7fc8bb8bbfb119d403</citedby><cites>FETCH-LOGICAL-c5760-7de6d90f4a9ed1eced487c03671bce98eb1128a2682b30f7fc8bb8bbfb119d403</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fcbic.201600320$$EPDF$$P50$$Gwiley$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fcbic.201600320$$EHTML$$P50$$Gwiley$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27441384$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>He, Kaizhang</creatorcontrib><creatorcontrib>Chou, Eldon T.</creatorcontrib><creatorcontrib>Begay, Shawn</creatorcontrib><creatorcontrib>Anderson, Emily M.</creatorcontrib><creatorcontrib>van Brabant Smith, Anja</creatorcontrib><title>Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing</title><title>Chembiochem : a European journal of chemical biology</title><addtitle>ChemBioChem</addtitle><description>The CRISPR‐Cas9 gene editing system requires Cas9 endonuclease and guide RNAs (either the natural dual RNA consisting of crRNA and tracrRNA or a chimeric single guide RNA) that direct site‐specific double‐stranded DNA cleavage. This communication describes a click ligation approach that uses alkyne–azide cycloaddition to generate a triazole‐linked single guide RNA (sgRNA). The conjugated sgRNA shows efficient and comparable genome editing activity to natural dual RNA and unmodified sgRNA constructs.
Editing efficiency: A triazole‐linked single guide RNA (sgRNA) was constructed by using click chemistry. The sgRNA showed efficient CRISPR‐Cas9 genome editing activity comparable to that of natural dual RNA and unmodified sgRNA constructs. These findings open new avenues for generating long sgRNA libraries in array or plate formats.</description><subject>Communication</subject><subject>Communications</subject><subject>conjugation</subject><subject>CRISPR-Cas Systems - genetics</subject><subject>CRISPR-Cas9</subject><subject>Editing</subject><subject>Gene Editing</subject><subject>gene technology</subject><subject>Molecular Structure</subject><subject>oligonucleotides</subject><subject>RNA, Guide, CRISPR-Cas Systems - chemistry</subject><subject>RNA, Guide, CRISPR-Cas Systems - metabolism</subject><subject>single guide RNA</subject><subject>Triazoles - chemistry</subject><subject>Triazoles - metabolism</subject><issn>1439-4227</issn><issn>1439-7633</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>EIF</sourceid><recordid>eNqNkc1v0zAYhyMEYmNw5YgsceGS4o8kti9IW1S6SqWMbggJDpbjvCnuUnvYyWD89aRKqQYXkCz563kf2e8vSZ4TPCEY09emsmZCMSkwZhQ_SI5JxmTKC8Ye7tcZpfwoeRLjBmMsC0YeJ0eUZxlhIjtOvpTebfq17qx3SLsaTW91249b36CrYPVP30K6sO4aanRp3boFNOttDWi1PEWND6hczS8vVmmpo0QzcICmte0G8GnyqNFthGf7-ST5-HZ6VZ6ni_ezeXm6SE3OC5zyGopa4ibTEmoCBupMcINZwUllQAqoCKFC00LQiuGGN0ZU1TCa4VzWGWYnyZvRe9NXW6gNuC7oVt0Eu9XhTnlt1Z83zn5Va3-r8qEfmGaD4NVeEPy3HmKntjYaaFvtwPdREUG5EFII8j9owWkmiRzQl3-hG98HN3RiR-WMcUF2wslImeBjDNAc3k2w2kWsdhGrQ8RDwYv7vz3gvzMdADkC320Ld__QqfJsXt6Xp2OtjR38ONTqcK0KzniuPi1n6vzzhwvB8ndqyX4B7RfA1Q</recordid><startdate>20161004</startdate><enddate>20161004</enddate><creator>He, Kaizhang</creator><creator>Chou, Eldon T.</creator><creator>Begay, Shawn</creator><creator>Anderson, Emily M.</creator><creator>van Brabant Smith, Anja</creator><general>Blackwell Publishing Ltd</general><general>Wiley Subscription Services, Inc</general><general>John Wiley and Sons Inc</general><scope>BSCLL</scope><scope>24P</scope><scope>WIN</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20161004</creationdate><title>Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing</title><author>He, Kaizhang ; Chou, Eldon T. ; Begay, Shawn ; Anderson, Emily M. ; van Brabant Smith, Anja</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5760-7de6d90f4a9ed1eced487c03671bce98eb1128a2682b30f7fc8bb8bbfb119d403</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Communication</topic><topic>Communications</topic><topic>conjugation</topic><topic>CRISPR-Cas Systems - genetics</topic><topic>CRISPR-Cas9</topic><topic>Editing</topic><topic>Gene Editing</topic><topic>gene technology</topic><topic>Molecular Structure</topic><topic>oligonucleotides</topic><topic>RNA, Guide, CRISPR-Cas Systems - chemistry</topic><topic>RNA, Guide, CRISPR-Cas Systems - metabolism</topic><topic>single guide RNA</topic><topic>Triazoles - chemistry</topic><topic>Triazoles - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>He, Kaizhang</creatorcontrib><creatorcontrib>Chou, Eldon T.</creatorcontrib><creatorcontrib>Begay, Shawn</creatorcontrib><creatorcontrib>Anderson, Emily M.</creatorcontrib><creatorcontrib>van Brabant Smith, Anja</creatorcontrib><collection>Istex</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Chembiochem : a European journal of chemical biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>He, Kaizhang</au><au>Chou, Eldon T.</au><au>Begay, Shawn</au><au>Anderson, Emily M.</au><au>van Brabant Smith, Anja</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing</atitle><jtitle>Chembiochem : a European journal of chemical biology</jtitle><addtitle>ChemBioChem</addtitle><date>2016-10-04</date><risdate>2016</risdate><volume>17</volume><issue>19</issue><spage>1809</spage><epage>1812</epage><pages>1809-1812</pages><issn>1439-4227</issn><eissn>1439-7633</eissn><abstract>The CRISPR‐Cas9 gene editing system requires Cas9 endonuclease and guide RNAs (either the natural dual RNA consisting of crRNA and tracrRNA or a chimeric single guide RNA) that direct site‐specific double‐stranded DNA cleavage. This communication describes a click ligation approach that uses alkyne–azide cycloaddition to generate a triazole‐linked single guide RNA (sgRNA). The conjugated sgRNA shows efficient and comparable genome editing activity to natural dual RNA and unmodified sgRNA constructs.
Editing efficiency: A triazole‐linked single guide RNA (sgRNA) was constructed by using click chemistry. The sgRNA showed efficient CRISPR‐Cas9 genome editing activity comparable to that of natural dual RNA and unmodified sgRNA constructs. These findings open new avenues for generating long sgRNA libraries in array or plate formats.</abstract><cop>Germany</cop><pub>Blackwell Publishing Ltd</pub><pmid>27441384</pmid><doi>10.1002/cbic.201600320</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Communication Communications conjugation CRISPR-Cas Systems - genetics CRISPR-Cas9 Editing Gene Editing gene technology Molecular Structure oligonucleotides RNA, Guide, CRISPR-Cas Systems - chemistry RNA, Guide, CRISPR-Cas Systems - metabolism single guide RNA Triazoles - chemistry Triazoles - metabolism |
| title | Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing |
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