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Nucleic Acids Research Advance Access originally published online on June 27, 2008
Nucleic Acids Research 2008 36(14):e90; doi:10.1093/nar/gkn415
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Nucleic Acids Research, 2008, Vol. 36, No. 14 e90
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Light activation of transcription: photocaging of nucleotides for control over RNA polymerization

André Vidal Pinheiro1,2, Pedro Baptista2,* and João Carlos Lima1

1REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa and 2CIGMH, Departamento Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal

*To whom correspondence should be addressed. Tel./Fax: +351 21 2948530; Email: pmvb{at}fct-unl.pt Correspondence may also be addressed to J. C. Lima. Tel./Fax: +351 212 948385; Email: lima{at}dq.fct.unl.pt

Received May 27, 2008. Revised June 12, 2008. Accepted June 16, 2008.

We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is initiated. In order to provide full control over the process, conditions involved in substrate release, nucleotide availability after release and the effect of the released coumarin in RNA polymerization were assessed in further detail. Together, our data provide the first direct evidence of control over enzymatic polymerization of nucleic acids through light. This approach may provide researchers with a unique tool for the study of biological processes at a molecular level.


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