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DC Field | Value | Language |
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dc.contributor.author | DUTTA, SWAGATA | en_US |
dc.contributor.author | SRIVATSAN, SEERGAZHI G. | en_US |
dc.date.accessioned | 2024-07-29T11:31:10Z | |
dc.date.available | 2024-07-29T11:31:10Z | |
dc.date.issued | 2024-07 | en_US |
dc.identifier.citation | Chemistry—An Asian Journal. | en_US |
dc.identifier.issn | 1861-471X | en_US |
dc.identifier.issn | 1861-4728 | en_US |
dc.identifier.uri | https://doi.org/10.1002/asia.202400475 | en_US |
dc.identifier.uri | http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/9019 | |
dc.description.abstract | We report a systematic study on controlling the enzyme activity of a terminal uridylyl transferase (TUTase) called SpCID1, which provides methods to effect site-specific incorporation of a single modified nucleotide analog at the 3'-end of an RNA oligonucleotide (ON). Responsive heterocycle-modified fluorescent UTP probes that are useful in analyzing non-canonical nucleic acid structures and azide- and alkyne-modified UTP analogs that are compatible for chemoenzymatic functionalization were used as study systems. In the first strategy, we balanced the concentration of essential metal ion cofactors (Mg2+ and Mn2+ ions) to restrict the processivity of the enzyme, which gave a very good control on the incorporation of clickable nucleotide analogs. In the second approach, borate that complexes with 2' and 3' oxygen atoms of a ribose sugar was used as a reversibly binding chelator to block repeated addition of nucleotide analogs. Notably, in the presence of heterocycle-modified fluorescent UTPs, we obtained single-nucleotide incorporated RNA products in reasonable yields, while with clickable nucleotides yields were very good. Further, 3'-end azide- and alkyne-labeled RNA ONs were post-enzymatically functionalized by CuAAC and SPAAC reactions with fluorescent probes. These strategies broaden the scope of TUTase in site-specifically installing modifications of different types onto RNA for various applications. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Wiley | en_US |
dc.subject | Chemistry | en_US |
dc.subject | 2024 | en_US |
dc.subject | 2024-JUL-WEEK2 | en_US |
dc.subject | TOC-JUL-2024 | en_US |
dc.title | Enzymatic functionalization of RNA oligonucleotides by terminal uridylyl transferase using fluorescent and clickable nucleotide analogs | en_US |
dc.type | Article | en_US |
dc.contributor.department | Dept. of Chemistry | en_US |
dc.identifier.sourcetitle | Chemistry—An Asian Journal | en_US |
dc.publication.originofpublisher | Foreign | en_US |
Appears in Collections: | JOURNAL ARTICLES |
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