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

0567916 - ÚMG 2023 RIV CZ eng A - Abstract
Miklík, Dalibor - Slavková, Martina - Hejnar, Jiří
KNOCK-ING ON HETEROCHROMATIN: STABILITY OF GAMMARETROVIRAL EXPRESSION AFTER INTEGRATION RETARGETING.
Czech Chemical Society Symposium Series. Roč. 20, č. 6 (2022), č. článku P-63. ISSN 2336-7202.
[Annual meeting of the National Institute of Virology and Bacteriology (NIVB) /1./. 30.11.2022-02.12.2022, Kutná Hora]
R&D Projects: GA MŠMT(CZ) LX22NPO5103
Institutional support: RVO:68378050
Keywords : retrovirus vector * integration * heterochromatin
OECD category: Virology
http://www.ccsss.cz/index.php/ccsss/issue/view/37/67

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.
Permanent Link: https://hdl.handle.net/11104/0339181