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Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection
Author(s) -
Pan Qiuwei,
de Ruiter Petra E.,
von Eije Karin J.,
Smits Ron,
Kwekkeboom Jaap,
Tilanus Hugo W.,
Berkhout Ben,
Janssen Harry L.A.,
van der Laan Luc J.W.
Publication year - 2011
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2011.02.035
Subject(s) - small hairpin rna , rna interference , microrna , biology , viral replication , hepatitis c virus , virology , interferon , rna , small interfering rna , virus , gene , genetics
RNA interference (RNAi) is widely used as a screening tool for the identification of host genes involved in viral infection. Due to the limitation of raw small interfering RNA (siRNA), we tested two commonly used short hairpin RNA (shRNA) lentiviral libraries to identify host factors involved in hepatitis C virus (HCV) infection. It was found that these shRNA library vectors caused non‐specific disturbance of HCV replication that was not due to toxicity or interferon response, but related to the high shRNA levels disturbing the endogenous microRNA biogenesis. The high shRNA levels achieved with these vectors reduced the levels of mature microRNAs, including miR‐122 known to promote HCV replication. Our findings extend the caution of potential off‐target effects of lentiviral shRNA libraries which appear unsuitable to screen microRNA regulated phenotypes, such as HCV replication.