Four base recognition by triplex-forming oligonucleotides at physiological pH
Four base recognition by triplex-forming oligonucleotides at physiological pH
We have achieved recognition of all 4 bp by triple helix formation at physiological pH, using triplex-forming oligonucleotides that contain four different synthetic nucleotides. BAU [20-aminoethoxy-5-(3-aminoprop-1-ynyl)uridine] recognizes AT base pairs with high affinity, MeP (3-methyl-2 aminopyridine) binds to GC at higher pHs than cytosine, while APP (6-(3-aminopropyl)-7-methyl-3H-pyrrolo[2,3-d]pyrimidin-2(7H)-one) and S [N-(4-(3-acetamidophenyl)thiazol-2-yl-acetamide)] bind to CG and TA base pairs, respectively.
Fluorescence melting and DNase I footprinting demonstrate successful triplex formation at a 19mer oligopurine sequence that contains two CG and two TA interruptions. The complexes are pH dependent, but are still stable at pH 7.0. BAU, MeP and APP retain considerable selectivity, and single base pair changes opposite these residues cause a large reduction in affinity. In contrast, S is less selective and tolerates CG pairs as well as TA.
helix formation, selective recognition, nucleoside analogs, stable triplexes, dual recognition, dna triplexes, binding motif, duplex dna, pairs, gene
3025-3032
Rusling, David A.
d397afa8-d5c0-49e7-abe0-401a985278c2
Powers, Vicki E.C.
0646d625-5039-454c-a8ad-0da6bc72ae8a
Ranasinghe, Rohan T.
b29fc8b4-2a66-430a-85fa-ff1c9c261f32
Wang, Yang
7bfb9a35-82f9-4580-a448-c4fbffc7959c
Osborne, Sadie D.
9777c83c-8998-4fd6-913e-04d3902ee9cd
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
23 May 2005
Rusling, David A.
d397afa8-d5c0-49e7-abe0-401a985278c2
Powers, Vicki E.C.
0646d625-5039-454c-a8ad-0da6bc72ae8a
Ranasinghe, Rohan T.
b29fc8b4-2a66-430a-85fa-ff1c9c261f32
Wang, Yang
7bfb9a35-82f9-4580-a448-c4fbffc7959c
Osborne, Sadie D.
9777c83c-8998-4fd6-913e-04d3902ee9cd
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Rusling, David A., Powers, Vicki E.C., Ranasinghe, Rohan T., Wang, Yang, Osborne, Sadie D., Brown, Tom and Fox, Keith R.
(2005)
Four base recognition by triplex-forming oligonucleotides at physiological pH.
Nucleic Acids Research, 33 (9), .
(doi:10.1093/nar/gki625).
Abstract
We have achieved recognition of all 4 bp by triple helix formation at physiological pH, using triplex-forming oligonucleotides that contain four different synthetic nucleotides. BAU [20-aminoethoxy-5-(3-aminoprop-1-ynyl)uridine] recognizes AT base pairs with high affinity, MeP (3-methyl-2 aminopyridine) binds to GC at higher pHs than cytosine, while APP (6-(3-aminopropyl)-7-methyl-3H-pyrrolo[2,3-d]pyrimidin-2(7H)-one) and S [N-(4-(3-acetamidophenyl)thiazol-2-yl-acetamide)] bind to CG and TA base pairs, respectively.
Fluorescence melting and DNase I footprinting demonstrate successful triplex formation at a 19mer oligopurine sequence that contains two CG and two TA interruptions. The complexes are pH dependent, but are still stable at pH 7.0. BAU, MeP and APP retain considerable selectivity, and single base pair changes opposite these residues cause a large reduction in affinity. In contrast, S is less selective and tolerates CG pairs as well as TA.
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Submitted date: 6 April 2005
Published date: 23 May 2005
Keywords:
helix formation, selective recognition, nucleoside analogs, stable triplexes, dual recognition, dna triplexes, binding motif, duplex dna, pairs, gene
Identifiers
Local EPrints ID: 24235
URI: http://eprints.soton.ac.uk/id/eprint/24235
ISSN: 0305-1048
PURE UUID: 31093f9c-361a-41f0-96e1-bea6780f7034
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Date deposited: 28 Mar 2006
Last modified: 16 Mar 2024 02:36
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Contributors
Author:
David A. Rusling
Author:
Vicki E.C. Powers
Author:
Rohan T. Ranasinghe
Author:
Yang Wang
Author:
Sadie D. Osborne
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