Cutting-edge
nanoelectrokinetic technology in this work
provides
a breakthrough for the present clinical demands of molecular diagnosis
to detect a trace amount of oncogenic mutation of DNA in a short time
without an erroneous PCR procedure. In this work, we combined the
sequence-specific labeling scheme of CRISPR/dCas9 and ion concentration
polarization (ICP) mechanism to separately preconcentrate target DNA
molecules for rapid detection. Using the mobility shift caused by
dCas9’s specific binding to the mutant, the mutated DNA and
normal DNA were distinguished in the microchip. Based on this technique,
we successfully demonstrated the dCas9-mediated 1-min detection of
single base substitution (SBS) in EGFR DNA, a carcinogenesis indicator.
Moreover, the presence/absence of target DNA was identified at a glance
like a commercial pregnancy test kit (two lines for positive and one
line for negative) by the distinct preconcentration mechanisms of
ICP, even at the 0.1% concentration of the target mutant
