Сборник содержит материалы научной мультиконференции «Биотех- нология медицине будущего»

Excimer forming pyrene probes for detection of RNA and point mutations in RNA 
Semikolenova O.A.
1
, Kim B.H.
2
, Venyaminova A.G.
3
, Novopashina D.S.
1,3
1
Novosibirsk State University, Novosibirsk, Russia 
2
 pohang University of Science and Technology (pOSTECH), pohang , Korea 
3
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk, Russia
Design of new fluorescent probes for nucleic acids detection and point mutations deter-
mination is the actual direction of modern bioorganic chemistry and molecular biology.
In spite of the serious progress in the design of fluorescent probes there are no probes satisfy 
for the complex of all necessary requirements. The ideal fluorescent probe must be highly 
stable for exo- and endo-nucleases digestion, has a good affinity to NA targets and consid-
erable sensitivity of fluorescence upon binding with target. Pyrene modified oligonucleotide 
probes capable to form excimers are prospective instruments for RNA and DNA detection due 
to their high sensitivity to the microenviroment.
The aim of our work is the design of excimer forming oligonucleotide probes on the basis 
of pyrene bearing oligo(2ʹ-O-methylribonucleotides) and demonstration of their applicability 
for RNA detection and point mutation determination.
MicroRNAs particularly microRNA let-7 are considered as diagnostics biomarkers and 
are the objects of investigation upon creation of new improved systems of RNA detection. The 
series of 5ʹ-bispyrene probes and tandem probes bearing two pyrene residues on the compo-
nents junction were synthesized using two different methods. The first method is a newly 
developed solid-phase synthesis based on the 5ʹ-hydroxyl group of immobilized oligo(2ʹ-О-
methylribonucleotide) activation by N,Nʹ-disuccinimidyl carbonate and subsequent inter-
action with pyrenemethylamine. The second method is based on the activation of 5ʹ-phosphate 
of deprotected oligo(2ʹ-O-methylribonucleotide) in solution followed by interaction with 
pyrenemethylamine. The most sensitive fluorescent probes were revealed using the compar-
ative study of their fluorescent properties upon hybridization with synthetic microRNA let-7a 
target. The possibility of point mutation detection using 5ʹ-bispyrene probes was demonstrated 
using fully matched RNA and mismatched RNA containing one point mutation. The strong 
dependence of sensitivity of point mutation detection on nucleotide context was demonstrated. 
Proposed excimer forming pyrene modified oligo(2ʹ-O-methylribonucleotides) can by 
efficiently used for RNAs detection and point mutations determination. 
The work was partially supported by Russian State funded budget project of ICBFM SB RAS
# АААА-А17-117020210021-7.

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Всероссийская мультиконференция с международным участием «Биотехнология – медицине будущего»
29 июня - 2 июля 2019 г., г. Новосибирск, Россия
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