Amplification and Re-Generation of LNA-Modified Libraries

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Standard

Amplification and Re-Generation of LNA-Modified Libraries. / Doessing, Holger; Hansen, Lykke H.; Veedu, Rakesh N.; Wengel, Jesper; Vester, Birte.

In: Molecules, Vol. 17, No. 11, 2012, p. 13087.

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Harvard

Doessing, H, Hansen, LH, Veedu, RN, Wengel, J & Vester, B 2012, 'Amplification and Re-Generation of LNA-Modified Libraries' Molecules, vol 17, no. 11, pp. 13087., 10.3390/molecules171113087

APA

Doessing, H., Hansen, L. H., Veedu, R. N., Wengel, J., & Vester, B. (2012). Amplification and Re-Generation of LNA-Modified Libraries. Molecules, 17(11), 13087. 10.3390/molecules171113087

CBE

Doessing H, Hansen LH, Veedu RN, Wengel J, Vester B. 2012. Amplification and Re-Generation of LNA-Modified Libraries. Molecules. 17(11):13087. Available from: 10.3390/molecules171113087

MLA

Vancouver

Doessing H, Hansen LH, Veedu RN, Wengel J, Vester B. Amplification and Re-Generation of LNA-Modified Libraries. Molecules. 2012;17(11):13087. Available from: 10.3390/molecules171113087

Author

Doessing, Holger; Hansen, Lykke H.; Veedu, Rakesh N.; Wengel, Jesper; Vester, Birte / Amplification and Re-Generation of LNA-Modified Libraries.

In: Molecules, Vol. 17, No. 11, 2012, p. 13087.

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Bibtex

@article{855459ac3b2045ba8d1a61baaeada3c0,
title = "Amplification and Re-Generation of LNA-Modified Libraries",
publisher = "Molecular Diversity Preservation International (M D P I)",
author = "Holger Doessing and Hansen, {Lykke H.} and Veedu, {Rakesh N.} and Jesper Wengel and Birte Vester",
year = "2012",
doi = "10.3390/molecules171113087",
volume = "17",
number = "11",
pages = "13087",
journal = "Molecules",
issn = "1420-3049",

}

RIS

TY - JOUR

T1 - Amplification and Re-Generation of LNA-Modified Libraries

A1 - Doessing,Holger

A1 - Hansen,Lykke H.

A1 - Veedu,Rakesh N.

A1 - Wengel,Jesper

A1 - Vester,Birte

AU - Doessing,Holger

AU - Hansen,Lykke H.

AU - Veedu,Rakesh N.

AU - Wengel,Jesper

AU - Vester,Birte

PB - Molecular Diversity Preservation International (M D P I)

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

KW - Chemistry

U2 - 10.3390/molecules171113087

DO - 10.3390/molecules171113087

JO - Molecules

JF - Molecules

SN - 1420-3049

IS - 11

VL - 17

SP - 13087

ER -