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DNA Hybridization in Thermoresponsive Polymer Nanoparticles
Author(s) -
Moura Leila M.,
Martinho Jose M. G.,
Farinha Jose Paulo S.
Publication year - 2010
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201000015
Subject(s) - förster resonance energy transfer , nanoparticle , polymer , ionic bonding , dna , poly(n isopropylacrylamide) , fluorescence , particle (ecology) , materials science , ionic strength , dna–dna hybridization , chemical engineering , chemistry , polymer chemistry , biophysics , nanotechnology , copolymer , organic chemistry , aqueous solution , ion , biochemistry , biology , physics , quantum mechanics , oceanography , geology , engineering
We achieve very high hybridization efficiencies by using a new method to immobilize DNA strands on the surface of thermoresponsive polymer nanoparticles. Hybridization efficiencies of about 70 % are obtained between the DNA immobilized in the particles and a complementary strand in solution, even at very low ionic strengths (1 m M ). The polymer nanoparticles have a glassy poly(methylmethacrylate) (PMMA) core and a thermoresponsive shell of poly( N ‐isopropylacrylamide) (PNIPAM) containing positive charges. After a DNA strand labeled with a fluorescence probe is loaded onto the particles at room temperature, the temperature is increased above the volume phase transition temperature of the PNIPAM shell, TVPT ≈28 °C. The collapse of the particle shell immobilizes the DNA while maintaining its availability for hybridization with a complementary strand. Förster resonance energy transfer (FRET) is used to detect the hybridization with a complementary DNA strand labeled with a FRET acceptor probe.