The added sequence will bind up the messenger RNA with the same powerful bonding that holds together a DNA double helix, thus creating a freak double-stranded RNA or a DNA-RNA hybrid. In such a state, the messenger RNA can no longer perform its job of passing on the genetic code.
"We have seen reductions of the specific product proteins as well as observed the expected DNA-RNA hybrids to show how the antisense is working," says Eric Wickstrom of the University of South Florida in Tampa.
Such a DNA-RNA hybrid
would be resistant to both RNase and DNase activity.
Another point that has to be taken into account is that the used antibody can react with different DNA-RNA hybrids
with different affinity (24), influencing the sensitivity of the assay for this miRNA and requiring additional calibration.
The H stands for hybrid, and this enzymatic activity degrades away the RNA pair of annealed DNA-RNA hybrids
. The product of this step is therefore a single-stranded DNA complement copy of the original RNA target.
High resolution of DNA-RNA hybrids
in situ by indirect immunofluorescence.
In each case, specific HPV DNA-RNA hybrids
Specificity, however, may be further improved because biotinylated oligonucleotides specific for HPV DNA sequences are used to capture HPV DNA-RNA hybrids
(instead of anti-DNA-RNA polyclonal antibody used previously) onto streptavidine-coated wells of a microtiter plate.