Colorimetric detection of eukaryotic gene expression with DNA-derivatized gold nanoparticles.
Thiol-linked DNA-gold nanoparticles were used in a novel colorimetric method to detect the presence of specific mRNA from a total RNA extract of yeast cells. The method allowed detection of expression of the FSY1 gene that encodes a specific fructose/H+ symporter in Saccharomyces bayanus PYCC 4565. FSY1 is strongly expressed when the yeast is grown in fructose as the sole carbon source, while cells cultivated in glucose as the sole carbon source repress gene expression. The presence of FSY1 mRNA is detected based on color change of a sample containing total RNA extracted from the organism and gold nanoparticles derivatized with a 15-mer of complementary single stranded DNA upon addition of NaCl. If FSY1 mRNA is present, the solution remains pink, changing to blue-purple in the absence of FSY1 mRNA. Direct detection of specific expression was possible from only 0.3 microg of unamplified total RNA without any further enhancement. This novel method is inexpensive, very easy to perform as no amplification or signal enhancement steps are necessary and takes less than 15 min to develop after total RNA extraction. No temperature control is necessary and color change can be easily detected visually.
Baptista
Pedro
P
CIGMH/SABT, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
Doria
Gonçalo
G
Henriques
David
D
Pereira
Eulália
E
Franco
Ricardo
R
D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D026901:Membrane Transport Proteins / Q000235:genetics; D049329:Nanostructures / Q000737:chemistry*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012440:Saccharomyces / Q000235:genetics*
D015342:DNA Probes; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D004247:DNA