Efficient preparation and properties of mRNAs containing a fluorescent cap analog: Anthraniloyl-m7GpppG
A method has been developed for synthesising fluorescently labeled capped mRNA. The method incorporates a single fluorescent molecule as part of the 5′-mRNA or oligonucleotide cap site. The fluorescent molecule, Ant-m 7 GTP is specifically incorporated into the cap site to yield Ant-m 7 GpppG-capped...
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| Veröffentlicht in: | Translation (Austin, Tex.) Tex.), 2015-02, Vol.3 (1) |
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| creator | Gunawardana, Dilantha Domashevskiy, Artem V Gayler, Ken R Goss, Dixie J |
| description | A method has been developed for synthesising fluorescently labeled capped mRNA. The method incorporates a single fluorescent molecule as part of the 5′-mRNA or oligonucleotide cap site. The fluorescent molecule, Ant-m
7
GTP is specifically incorporated into the cap site to yield Ant-m
7
GpppG-capped mRNA or oligonucleotide. Efficient capping was observed with 60-100% of the RNA transcripts capped with the fluorescent molecule. The Ant-m
7
G derivative, which has been previously shown to interact with the eukaryotic cap binding protein eIF4E, is shown in this paper to be a substrate for the Vaccinia capping enzyme and the DCP2 decapping enzyme from Arabidopsis. Further, the Ant-m
7
GTP-capped RNA is readily translated. This Ant-m
7
GTP-capped RNA provides an important tool for monitoring capping reactions, translation, and biophysical studies. |
| doi_str_mv | 10.4161/21690731.2014.988538 |
| format | Article |
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7
GTP is specifically incorporated into the cap site to yield Ant-m
7
GpppG-capped mRNA or oligonucleotide. Efficient capping was observed with 60-100% of the RNA transcripts capped with the fluorescent molecule. The Ant-m
7
G derivative, which has been previously shown to interact with the eukaryotic cap binding protein eIF4E, is shown in this paper to be a substrate for the Vaccinia capping enzyme and the DCP2 decapping enzyme from Arabidopsis. Further, the Ant-m
7
GTP-capped RNA is readily translated. This Ant-m
7
GTP-capped RNA provides an important tool for monitoring capping reactions, translation, and biophysical studies.</description><identifier>EISSN: 2169-0731</identifier><identifier>DOI: 10.4161/21690731.2014.988538</identifier><identifier>PMID: 26779415</identifier><language>eng</language><publisher>Taylor & Francis</publisher><subject>Ant, Anthraniloyl ; Ant-GTP, Anthraniloyl guanosine triphosphate ; Ant-m ; capped mRNA ; decapping assay ; Fluorescence ; GMP, Anthraniloyl-7-methyl guanosine monophosphate ; GTP, Anthraniloyl-7-methyl guanosine triphosphate ; imaging ; IVT, In vitro translation ; Methods ; OMP, Outer Membrane Protein ; RNA capping ; SAM, S-Adenosyl Methionine</subject><ispartof>Translation (Austin, Tex.), 2015-02, Vol.3 (1)</ispartof><rights>Taylor & Francis Group, LLC 2015</rights><rights>Taylor & Francis Group, LLC 2015 Taylor & Francis Group</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682804/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682804/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,724,777,781,882,27905,27906,53772,53774</link.rule.ids></links><search><creatorcontrib>Gunawardana, Dilantha</creatorcontrib><creatorcontrib>Domashevskiy, Artem V</creatorcontrib><creatorcontrib>Gayler, Ken R</creatorcontrib><creatorcontrib>Goss, Dixie J</creatorcontrib><title>Efficient preparation and properties of mRNAs containing a fluorescent cap analog: Anthraniloyl-m7GpppG</title><title>Translation (Austin, Tex.)</title><description>A method has been developed for synthesising fluorescently labeled capped mRNA. The method incorporates a single fluorescent molecule as part of the 5′-mRNA or oligonucleotide cap site. The fluorescent molecule, Ant-m
7
GTP is specifically incorporated into the cap site to yield Ant-m
7
GpppG-capped mRNA or oligonucleotide. Efficient capping was observed with 60-100% of the RNA transcripts capped with the fluorescent molecule. The Ant-m
7
G derivative, which has been previously shown to interact with the eukaryotic cap binding protein eIF4E, is shown in this paper to be a substrate for the Vaccinia capping enzyme and the DCP2 decapping enzyme from Arabidopsis. Further, the Ant-m
7
GTP-capped RNA is readily translated. This Ant-m
7
GTP-capped RNA provides an important tool for monitoring capping reactions, translation, and biophysical studies.</description><subject>Ant, Anthraniloyl</subject><subject>Ant-GTP, Anthraniloyl guanosine triphosphate</subject><subject>Ant-m</subject><subject>capped mRNA</subject><subject>decapping assay</subject><subject>Fluorescence</subject><subject>GMP, Anthraniloyl-7-methyl guanosine monophosphate</subject><subject>GTP, Anthraniloyl-7-methyl guanosine triphosphate</subject><subject>imaging</subject><subject>IVT, In vitro translation</subject><subject>Methods</subject><subject>OMP, Outer Membrane Protein</subject><subject>RNA capping</subject><subject>SAM, S-Adenosyl Methionine</subject><issn>2169-0731</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNpVkFFLwzAQx4Mgbsx9Ax_6BTqTNmlaH5Qx5hSGguhzuKRJF2mTkHbKvr0t0wfv5bj_3f25-yF0Q_CKkoLcZqSoMM_JKsOErqqyZHl5geaTnE76DC37_hOPUXFMM3qFZlnBeUUJm6Nma4xVVrshCVEHiDBY7xJw9Vj7oONgdZ94k3RvL-s-Ud4NYJ11TQKJaY8-6l5NywrCuAStb-6StRsOEZxt_alNO74LIeyu0aWBttfL37xAH4_b981Tun_dPW_W-9QSxsuUSyMpLrA2UnGVS2KkLpniOWOgGYWCEKq0wRIKzTnB4xSTRpNKMlJzXeYLdH_2DUfZ6Xq6LUIrQrQdxJPwYMX_jrMH0fgvQYsyKzEdDR7OBtYZHzv49rGtxQCn1kczfqVsL3KCxYRe_KEXE3pxRp__AE4Gez8</recordid><startdate>20150202</startdate><enddate>20150202</enddate><creator>Gunawardana, Dilantha</creator><creator>Domashevskiy, Artem V</creator><creator>Gayler, Ken R</creator><creator>Goss, Dixie J</creator><general>Taylor & Francis</general><scope>5PM</scope></search><sort><creationdate>20150202</creationdate><title>Efficient preparation and properties of mRNAs containing a fluorescent cap analog: Anthraniloyl-m7GpppG</title><author>Gunawardana, Dilantha ; Domashevskiy, Artem V ; Gayler, Ken R ; Goss, Dixie J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-i1578-7bfb4060efbc7c3b1fbe85c7355ae54a6114cef0ba6e7710bc75bfe19b51d7e83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Ant, Anthraniloyl</topic><topic>Ant-GTP, Anthraniloyl guanosine triphosphate</topic><topic>Ant-m</topic><topic>capped mRNA</topic><topic>decapping assay</topic><topic>Fluorescence</topic><topic>GMP, Anthraniloyl-7-methyl guanosine monophosphate</topic><topic>GTP, Anthraniloyl-7-methyl guanosine triphosphate</topic><topic>imaging</topic><topic>IVT, In vitro translation</topic><topic>Methods</topic><topic>OMP, Outer Membrane Protein</topic><topic>RNA capping</topic><topic>SAM, S-Adenosyl Methionine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gunawardana, Dilantha</creatorcontrib><creatorcontrib>Domashevskiy, Artem V</creatorcontrib><creatorcontrib>Gayler, Ken R</creatorcontrib><creatorcontrib>Goss, Dixie J</creatorcontrib><collection>PubMed Central (Full Participant titles)</collection><jtitle>Translation (Austin, Tex.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gunawardana, Dilantha</au><au>Domashevskiy, Artem V</au><au>Gayler, Ken R</au><au>Goss, Dixie J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient preparation and properties of mRNAs containing a fluorescent cap analog: Anthraniloyl-m7GpppG</atitle><jtitle>Translation (Austin, Tex.)</jtitle><date>2015-02-02</date><risdate>2015</risdate><volume>3</volume><issue>1</issue><eissn>2169-0731</eissn><abstract>A method has been developed for synthesising fluorescently labeled capped mRNA. The method incorporates a single fluorescent molecule as part of the 5′-mRNA or oligonucleotide cap site. The fluorescent molecule, Ant-m
7
GTP is specifically incorporated into the cap site to yield Ant-m
7
GpppG-capped mRNA or oligonucleotide. Efficient capping was observed with 60-100% of the RNA transcripts capped with the fluorescent molecule. The Ant-m
7
G derivative, which has been previously shown to interact with the eukaryotic cap binding protein eIF4E, is shown in this paper to be a substrate for the Vaccinia capping enzyme and the DCP2 decapping enzyme from Arabidopsis. Further, the Ant-m
7
GTP-capped RNA is readily translated. This Ant-m
7
GTP-capped RNA provides an important tool for monitoring capping reactions, translation, and biophysical studies.</abstract><pub>Taylor & Francis</pub><pmid>26779415</pmid><doi>10.4161/21690731.2014.988538</doi><oa>free_for_read</oa></addata></record> |
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| identifier | EISSN: 2169-0731 |
| ispartof | Translation (Austin, Tex.), 2015-02, Vol.3 (1) |
| issn | 2169-0731 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4682804 |
| source | EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Ant, Anthraniloyl Ant-GTP, Anthraniloyl guanosine triphosphate Ant-m capped mRNA decapping assay Fluorescence GMP, Anthraniloyl-7-methyl guanosine monophosphate GTP, Anthraniloyl-7-methyl guanosine triphosphate imaging IVT, In vitro translation Methods OMP, Outer Membrane Protein RNA capping SAM, S-Adenosyl Methionine |
| title | Efficient preparation and properties of mRNAs containing a fluorescent cap analog: Anthraniloyl-m7GpppG |
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