KNOCK-ING ON HETEROCHROMATIN: STABILITY OF GAMMARETROVIRAL EXPRESSION AFTER INTEGRATION RETARGETING

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68378050%3A_____%2F22%3A00567916" target="_blank" >RIV/68378050:_____/22:00567916 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.ccsss.cz/index.php/ccsss/issue/view/37/67" target="_blank" >http://www.ccsss.cz/index.php/ccsss/issue/view/37/67</a>

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

KNOCK-ING ON HETEROCHROMATIN: STABILITY OF GAMMARETROVIRAL EXPRESSION AFTER INTEGRATION RETARGETING

Popis výsledku v původním jazyce

Retroviruses integrate their genome into the genome of the infected cell. Integrated proviruses form an inseparable part of the host genome and express viral genes effectively. Epigenetic silencing may however disrupt the proviral expression. We have previously shown that retroviruses differ in sensitivity to epigenetic silencing and that the features at the integration site can affect the stability of proviral expression. Vectors derived from the murine leukemia virus (MLV) –widely used representative of the gammaretrovirus genus –performed as the most efficient retrovirus-derived vector in human cells. Here, we further investigate the gammaretroviral expression in human cells.We constructed retroviral vectors from diverse gammaretroviruses and transduced the human K562 cell line. We further tested if disruption of natural promoter/enhancer targeting affects the gammaretroviral expression. Our results indicate that the vectors express transduced genes stably in time irrespective of integration preference. We further corroborated the gammaretroviral expression stability in heterochromatin by CRISPR-directed knock-in into predicted lamin-associated domains.Our results demonstrate the general ability of gammaretroviruses to stably express transduced genes in human cells. Moreover, we show that other-than-MLV gammaretroviruses may be used for the construction of heterochromatin-resistant expression vectors for transgenesis.

Název v anglickém jazyce

KNOCK-ING ON HETEROCHROMATIN: STABILITY OF GAMMARETROVIRAL EXPRESSION AFTER INTEGRATION RETARGETING

Popis výsledku anglicky

Retroviruses integrate their genome into the genome of the infected cell. Integrated proviruses form an inseparable part of the host genome and express viral genes effectively. Epigenetic silencing may however disrupt the proviral expression. We have previously shown that retroviruses differ in sensitivity to epigenetic silencing and that the features at the integration site can affect the stability of proviral expression. Vectors derived from the murine leukemia virus (MLV) –widely used representative of the gammaretrovirus genus –performed as the most efficient retrovirus-derived vector in human cells. Here, we further investigate the gammaretroviral expression in human cells.We constructed retroviral vectors from diverse gammaretroviruses and transduced the human K562 cell line. We further tested if disruption of natural promoter/enhancer targeting affects the gammaretroviral expression. Our results indicate that the vectors express transduced genes stably in time irrespective of integration preference. We further corroborated the gammaretroviral expression stability in heterochromatin by CRISPR-directed knock-in into predicted lamin-associated domains.Our results demonstrate the general ability of gammaretroviruses to stably express transduced genes in human cells. Moreover, we show that other-than-MLV gammaretroviruses may be used for the construction of heterochromatin-resistant expression vectors for transgenesis.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10607 - Virology

Projekt

<a href="/cs/project/LX22NPO5103" target="_blank" >LX22NPO5103: Národní institut virologie a bakteriologie</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2022

Kód důvěrnosti údajů

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