A system for in vitro selection of fully 2′-modified RNA aptamers
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1.
Brandeis University
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2.
California Institute of Technology
Abstract
SFM4-3, KOD DGLNK, and Therminator polymerase are investigated for their compatibility with SELection with Modified Aptamers (SELMA), an aptamer discovery method that enables incorporation of large nucleobase modifications such as glycans. We demonstrated that with suitable modifications to the primer design and protocol, these enzymes are compatible with SELMA, enabling 2′-fluoro or 2′-methoxy ribose modifications at all positions. In the case of 2′-fluoro modifications, Therminator exhibits cleaner incorporation of an alkyne-modified nucleobase for click chemistry.
Copyright and License (English)
© The Royal Society of Chemistry 2025. This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. You can use material from this article in other publications, without requesting further permission from the RSC, provided that the correct acknowledgement is given and it is not used for commercial purposes.
Acknowledgement (English)
We would like to thank Dr Floyd Romesburg for the plasmid of SFM4-3 and Dr Satoshi Obika for the plasmid of KOD DGLNK. Dr Richard Redman and Dr Lizbeth Hedstrom are gratefully acknowledged for helpful discussions.
Funding (English)
This work was supported by NIH Grants R01GM151492 (I. J. K.), R01GM127920 (L. C. H.-W. and I. J. K.) and T32GM139798 (K. B. F.) as well as the NSF Graduate Research Fellowship No. 2139752 (E. D. Z.). I. J. K. acknowledges the NIH Shared Instrumentation grant S10OD034395 for purchase of an NMR spectrometer.
Data Availability (English)
The derived data supporting these findings can be found in the ESI.†
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Additional details
- National Institutes of Health
- R01GM151492
- National Institutes of Health
- R01GM127920
- National Institutes of Health
- T32GM139798
- National Science Foundation
- Graduate Research Fellowship 2139752
- National Institutes of Health
- Shared Instrumentation grant S10OD034395
- Accepted
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2025-01-28Accepted
- Available
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2025-02-06First published
- Publication Status
- Published