Abstract

Spatial and temporal control of molecular mechanisms can be achieved using photolabile bonds that connect biomolecules to protective caging groups, which can be cleaved upon irradiation of a specific wavelength, releasing the biomolecule ready-to-use. Here we apply and improve a previously reported strategy to tightly control in vitro transcription reactions. The strategy involves two caging molecules that block both ATP and GTP nucleotides. Additionally, we designed a molecular beacon complementary to the synthesized mRNA to infer its presence through a light signal. Upon release of both nucleotides through a specific monochromatic light (390 and 325 nm) we attain a light signal indicative of a successful in vitro transcription reaction. Similarly, in the absence of irradiation, no intense fluorescence signal was obtained. We believe this strategy could further be applied to DNA synthesis or the development of logic gates.

Original languageEnglish
Pages (from-to)751-756
Number of pages6
JournalPhotochemical and Photobiological Sciences
Volume13
Issue number5
DOIs
Publication statusPublished - 2014

Keywords

  • MOLECULAR BEACONS
  • PHOTOCLEAVAGE
  • ESTERS
  • TOOLS
  • RNA

Cite this

Reimao-Pinto, Madalena M. ; Cordeiro, Ana ; Almeida, Carina ; Pinheiro, Andre V. ; Moro, Artur ; Lima, Joao C. ; Baptista, Pedro. / Dual-color control of nucleotide polymerization sensed by a fluorescence actuator. In: Photochemical and Photobiological Sciences. 2014 ; Vol. 13, No. 5. pp. 751-756.
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abstract = "Spatial and temporal control of molecular mechanisms can be achieved using photolabile bonds that connect biomolecules to protective caging groups, which can be cleaved upon irradiation of a specific wavelength, releasing the biomolecule ready-to-use. Here we apply and improve a previously reported strategy to tightly control in vitro transcription reactions. The strategy involves two caging molecules that block both ATP and GTP nucleotides. Additionally, we designed a molecular beacon complementary to the synthesized mRNA to infer its presence through a light signal. Upon release of both nucleotides through a specific monochromatic light (390 and 325 nm) we attain a light signal indicative of a successful in vitro transcription reaction. Similarly, in the absence of irradiation, no intense fluorescence signal was obtained. We believe this strategy could further be applied to DNA synthesis or the development of logic gates.",
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Dual-color control of nucleotide polymerization sensed by a fluorescence actuator. / Reimao-Pinto, Madalena M.; Cordeiro, Ana; Almeida, Carina; Pinheiro, Andre V.; Moro, Artur; Lima, Joao C.; Baptista, Pedro.

In: Photochemical and Photobiological Sciences, Vol. 13, No. 5, 2014, p. 751-756.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dual-color control of nucleotide polymerization sensed by a fluorescence actuator

AU - Reimao-Pinto, Madalena M.

AU - Cordeiro, Ana

AU - Almeida, Carina

AU - Pinheiro, Andre V.

AU - Moro, Artur

AU - Lima, Joao C.

AU - Baptista, Pedro

PY - 2014

Y1 - 2014

N2 - Spatial and temporal control of molecular mechanisms can be achieved using photolabile bonds that connect biomolecules to protective caging groups, which can be cleaved upon irradiation of a specific wavelength, releasing the biomolecule ready-to-use. Here we apply and improve a previously reported strategy to tightly control in vitro transcription reactions. The strategy involves two caging molecules that block both ATP and GTP nucleotides. Additionally, we designed a molecular beacon complementary to the synthesized mRNA to infer its presence through a light signal. Upon release of both nucleotides through a specific monochromatic light (390 and 325 nm) we attain a light signal indicative of a successful in vitro transcription reaction. Similarly, in the absence of irradiation, no intense fluorescence signal was obtained. We believe this strategy could further be applied to DNA synthesis or the development of logic gates.

AB - Spatial and temporal control of molecular mechanisms can be achieved using photolabile bonds that connect biomolecules to protective caging groups, which can be cleaved upon irradiation of a specific wavelength, releasing the biomolecule ready-to-use. Here we apply and improve a previously reported strategy to tightly control in vitro transcription reactions. The strategy involves two caging molecules that block both ATP and GTP nucleotides. Additionally, we designed a molecular beacon complementary to the synthesized mRNA to infer its presence through a light signal. Upon release of both nucleotides through a specific monochromatic light (390 and 325 nm) we attain a light signal indicative of a successful in vitro transcription reaction. Similarly, in the absence of irradiation, no intense fluorescence signal was obtained. We believe this strategy could further be applied to DNA synthesis or the development of logic gates.

KW - MOLECULAR BEACONS

KW - PHOTOCLEAVAGE

KW - ESTERS

KW - TOOLS

KW - RNA

U2 - 10.1039/c3pp50438g

DO - 10.1039/c3pp50438g

M3 - Article

VL - 13

SP - 751

EP - 756

JO - Photochemical and Photobiological Sciences

JF - Photochemical and Photobiological Sciences

SN - 1474-905X

IS - 5

ER -