Amplification and re-generation of LNA-modified libraries

Doessing, Holger, Hanse, Lykke H., Veedu, Rakesh N., Wengel, Jesper and Vester, Birte (2012) Amplification and re-generation of LNA-modified libraries. Molecules, 17 11: 13087-13097. doi:10.3390/molecules171113087


Author Doessing, Holger
Hanse, Lykke H.
Veedu, Rakesh N.
Wengel, Jesper
Vester, Birte
Title Amplification and re-generation of LNA-modified libraries
Journal name Molecules   Check publisher's open access policy
ISSN 1420-3049
Publication date 2012
Sub-type Article (original research)
DOI 10.3390/molecules171113087
Volume 17
Issue 11
Start page 13087
End page 13097
Total pages 11
Place of publication Basel, Switzerland
Publisher M D P I AG
Collection year 2013
Language eng
Abstract Locked nucleic acids (LNA) confer high thermal stability and nuclease resistance to oligonucleotides. The discovery of polymerases that accept LNA triphosphates has led us to propose a scheme for the amplification and re-generation of LNA-containing oligonucleotide libraries. Such libraries could be used for in vitro selection of e.g., native LNA aptamers. We maintained an oligonucleotide library encoding 40 randomized positions with LNA ATP, GTP, CTP, and TTP for 7 rounds of ‘mock’ in vitro selection in the absence of a target and analyzed the sequence composition after rounds 1, 4 and 7. We observed a decrease in LNA-A content from 20.5% in round 1 to 6.6% in round 7. This decrease was accompanied by a substantial bias against successive LNA-As (poly-LNA adenosine tracts) and a relative over-representation of single LNA-As. Maintaining a library with LNA TTP yielded similar results. Together, these results suggest that dispersed LNA monomers are tolerated in our in vitro selection protocol, and that LNA-modified libraries can be sustained for up to at least seven selection rounds, albeit at reduced levels. This enables the discovery of native LNA aptamers and similar oligonucleotide structures.
Keyword Locked nucleic acid (LNA)
In vitro selection
Systematic evolution of ligands by exponential enrichment (SELEX)
Aptamer
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Chemistry and Molecular Biosciences
 
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Created: Fri, 23 Nov 2012, 09:49:21 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences