Use of siRNAs to manage HBV model infections

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F19%3AN0000175" target="_blank" >RIV/00027162:_____/19:N0000175 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Use of siRNAs to manage HBV model infections

Popis výsledku v původním jazyce

2019 International HBV Meeting, Melbourne, 1.10.-5.10.2019 – Poster. Our current work is devoted to studying and optimizing the ability of siRNAs to silence HBV gene expression in HepG2.2.15 cells in vitro, The siRNAs 1407,1794, 263 and 3, all leading positive controls were delivered to HepG2.2.15 cells. Thereafter, RNAi effects were monitored with reference to hepatitis B surface antigen (HBsAg) gene expression as detected by quantitative RT-PCR (RT-qPCR) on the 2nd, 4th and 6th day after siRNA delivery. Various doses of siRNAs (1nmol, 20pmol, 2pmol and 0.2pmol) were used to study the silencing of HBsAg gene expression. We found that treatment of cells with 1nmol, 20pmol, or 2pmol of siRNA/well resulted in highly significant knockdown of HBsAg expression levels (%) on 2nd and 4th days post transfection compared to the situation with matched control siRNAs. However, on 6th day the siRNA-mediated knockdown of HBsAg expression decreased. At lower doses of siRNAs (0.2pmol/well), 1407 was able to modulate HBsAg gene expression on the 2nd day after transfection, unlike the remaining siRNAs. This effect did not persist into the 4th and 6th day after transfection. Among the four leading positive control siRNAs, 1407 appears the most potent at modulating HBsAg expression compared with the other three remaining positive control siRNAs. Differences in inhibition among various siRNAs may be attributed to differences in affinity between siRNA guide strands and siRNA binding proteins, together with variations in the natures of the secondary and/or tertiary structures in mRNAs targeted for hydrolytic cleavage. Our next steps are to look at how siRNA chemical modifications and different chemical entities might extend the durability of siRNA-mediated HBV gene silencing effects in vitro and then in vivo, paving the way for new siRNA therapeutic approaches for CHB treatment.

Název v anglickém jazyce

Use of siRNAs to manage HBV model infections

Popis výsledku anglicky

2019 International HBV Meeting, Melbourne, 1.10.-5.10.2019 – Poster. Our current work is devoted to studying and optimizing the ability of siRNAs to silence HBV gene expression in HepG2.2.15 cells in vitro, The siRNAs 1407,1794, 263 and 3, all leading positive controls were delivered to HepG2.2.15 cells. Thereafter, RNAi effects were monitored with reference to hepatitis B surface antigen (HBsAg) gene expression as detected by quantitative RT-PCR (RT-qPCR) on the 2nd, 4th and 6th day after siRNA delivery. Various doses of siRNAs (1nmol, 20pmol, 2pmol and 0.2pmol) were used to study the silencing of HBsAg gene expression. We found that treatment of cells with 1nmol, 20pmol, or 2pmol of siRNA/well resulted in highly significant knockdown of HBsAg expression levels (%) on 2nd and 4th days post transfection compared to the situation with matched control siRNAs. However, on 6th day the siRNA-mediated knockdown of HBsAg expression decreased. At lower doses of siRNAs (0.2pmol/well), 1407 was able to modulate HBsAg gene expression on the 2nd day after transfection, unlike the remaining siRNAs. This effect did not persist into the 4th and 6th day after transfection. Among the four leading positive control siRNAs, 1407 appears the most potent at modulating HBsAg expression compared with the other three remaining positive control siRNAs. Differences in inhibition among various siRNAs may be attributed to differences in affinity between siRNA guide strands and siRNA binding proteins, together with variations in the natures of the secondary and/or tertiary structures in mRNAs targeted for hydrolytic cleavage. Our next steps are to look at how siRNA chemical modifications and different chemical entities might extend the durability of siRNA-mediated HBV gene silencing effects in vitro and then in vivo, paving the way for new siRNA therapeutic approaches for CHB treatment.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10607 - Virology

Projekt

<a href="/cs/project/EF15_003%2F0000495" target="_blank" >EF15_003/0000495: FIT (Farmakologie, Imunoterapie, nanoToxikologie)</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů