TY - JOUR
T1 - Biophysical Characterization of Novel DNA Aptamers against K103N/Y181C Double Mutant HIV-1 Reverse Transcriptase
AU - Ratanabunyong, Siriluk
AU - Seetaha, Supaphorn
AU - Hannongbua, Supa
AU - Yanaka, Saeko
AU - Yagi-Utsumi, Maho
AU - Kato, Koichi
AU - Paemanee, Atchara
AU - Choowongkomon, Kiattawee
N1 - Funding Information:
This research was funded by Kasetsart University Research and Development Institute, KURDI (FF (KU) 17.64), the National Research Council of Thailand (NRCT), the National Nanotech-nology Center (NANOTEC), NSTDA, The Ministry of Higher Education, Science, Research and In-novation of Thailand, through its program of Research Network of NANOTEC (RNN), the Thailand Research Fund (DBG5980001) and Joint Research by Exploratory Research Center on Life and Living Systems (ExCELLS) (ExCELLS program No, 19-310), and Nanotechnology Platform Program (Mol-ecule and Material Synthesis) of MEXT, Japan for the NMR experiments. The APC was fund by the Faculty of Science, Kasetsart University.
Funding Information:
Funding: This research was funded by Kasetsart University Research and Development Institute, KURDI (FF (KU) 17.64), the National Research Council of Thailand (NRCT), the National Nanotechnology Center (NANOTEC), NSTDA, The Ministry of Higher Education, Science, Research and Innovation of Thailand, through its program of Research Network of NANOTEC (RNN), the Thailand Research Fund (DBG5980001) and Joint Research by Exploratory Research Center on Life and Living Systems (ExCELLS) (ExCELLS program No, 19-310), and Nanotechnology Platform Program (Molecule and Material Synthesis) of MEXT, Japan for the NMR experiments. The APC was fund by the Faculty of Science, Kasetsart University.
Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/1/1
Y1 - 2022/1/1
N2 - The human immunodeficiency virus type-1 Reverse Transcriptase (HIV-1 RT) plays a piv-otal role in essential viral replication and is the main target for antiviral therapy. The anti-HIV-1 RT drugs address resistance-associated mutations. This research focused on isolating the potential specific DNA aptamers against K103N/Y181C double mutant HIV-1 RT. Five DNA aptamers showed low IC50 values against both the KY-mutant HIV-1 RT and wildtype (WT) HIV-1 RT. The kinetic binding affinity forms surface plasmon resonance of both KY-mutant and WT HIV-1 RTs in the range of 0.06–2 μM and 0.15–2 μM, respectively. Among these aptamers, the KY44 aptamer was chosen to study the interaction of HIV-1 RTs-DNA aptamer complex by NMR experiments. The NMR results indicate that the aptamer could interact with both WT and KY-mutant HIV-1 RT at the NNRTI drug binding pocket by inducing a chemical shift at methionine residues. Furthermore, KY44 could inhibit pseudo-HIV particle infection in HEK293 cells with nearly 80% inhibition and showed low cytotoxicity on HEK293 cells. These together indicated that the KY44 aptamer could be a potential inhibitor of both WT and KY-mutant HIV-RT.
AB - The human immunodeficiency virus type-1 Reverse Transcriptase (HIV-1 RT) plays a piv-otal role in essential viral replication and is the main target for antiviral therapy. The anti-HIV-1 RT drugs address resistance-associated mutations. This research focused on isolating the potential specific DNA aptamers against K103N/Y181C double mutant HIV-1 RT. Five DNA aptamers showed low IC50 values against both the KY-mutant HIV-1 RT and wildtype (WT) HIV-1 RT. The kinetic binding affinity forms surface plasmon resonance of both KY-mutant and WT HIV-1 RTs in the range of 0.06–2 μM and 0.15–2 μM, respectively. Among these aptamers, the KY44 aptamer was chosen to study the interaction of HIV-1 RTs-DNA aptamer complex by NMR experiments. The NMR results indicate that the aptamer could interact with both WT and KY-mutant HIV-1 RT at the NNRTI drug binding pocket by inducing a chemical shift at methionine residues. Furthermore, KY44 could inhibit pseudo-HIV particle infection in HEK293 cells with nearly 80% inhibition and showed low cytotoxicity on HEK293 cells. These together indicated that the KY44 aptamer could be a potential inhibitor of both WT and KY-mutant HIV-RT.
UR - http://www.scopus.com/inward/record.url?scp=85122152160&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85122152160&partnerID=8YFLogxK
U2 - 10.3390/molecules27010285
DO - 10.3390/molecules27010285
M3 - Article
C2 - 35011517
AN - SCOPUS:85122152160
SN - 1420-3049
VL - 27
JO - Molecules
JF - Molecules
IS - 1
M1 - 285
ER -
