Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection

Qiuwei Pan; Petra E. de Ruiter; Karin J. Von Eije; Ron Smits; Jaap Kwekkeboom; Hugo W. Tilanus; Ben Berkhout; Harry L. A. Janssen; Luc J. W. van der Laan

doi:10.1016/j.febslet.2011.02.035

Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection

Qiuwei Pan
, Petra E. de Ruiter
, Karin J. Von Eije
, Ron Smits
, Jaap Kwekkeboom
, Hugo W. Tilanus
, Ben Berkhout
, Harry L. A. Janssen
, Luc J. W. van der Laan

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved

Original language	English
Pages (from-to)	1025-1030
Journal	FEBS letters
Volume	585
Issue number	7
DOIs	https://doi.org/10.1016/j.febslet.2011.02.035
Publication status	Published - 2011

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SDG 3 Good Health and Well-being

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Pan, Q., de Ruiter, P. E., Von Eije, K. J., Smits, R., Kwekkeboom, J., Tilanus, H. W., Berkhout, B., Janssen, H. L. A., & van der Laan, L. J. W. (2011). Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection. FEBS letters, 585(7), 1025-1030. https://doi.org/10.1016/j.febslet.2011.02.035

@article{f73f7983dec7409b9ed96ae54d5cbafc,

title = "Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection",

abstract = "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. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved",

author = "Qiuwei Pan and \{de Ruiter\}, \{Petra E.\} and \{Von Eije\}, \{Karin J.\} and Ron Smits and Jaap Kwekkeboom and Tilanus, \{Hugo W.\} and Ben Berkhout and Janssen, \{Harry L. A.\} and \{van der Laan\}, \{Luc J. W.\}",

year = "2011",

doi = "10.1016/j.febslet.2011.02.035",

language = "English",

volume = "585",

pages = "1025--1030",

journal = "FEBS letters",

issn = "0014-5793",

publisher = "Wiley Blackwell",

number = "7",

}

Pan, Q, de Ruiter, PE, Von Eije, KJ, Smits, R, Kwekkeboom, J, Tilanus, HW, Berkhout, B, Janssen, HLA & van der Laan, LJW 2011, 'Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection', FEBS letters, vol. 585, no. 7, pp. 1025-1030. https://doi.org/10.1016/j.febslet.2011.02.035

TY - JOUR

T1 - Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection

AU - Pan, Qiuwei

AU - de Ruiter, Petra E.

AU - Von Eije, Karin J.

AU - Smits, Ron

AU - Kwekkeboom, Jaap

AU - Tilanus, Hugo W.

AU - Berkhout, Ben

AU - Janssen, Harry L. A.

AU - van der Laan, Luc J. W.

PY - 2011

Y1 - 2011

N2 - 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. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved

AB - 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. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved

U2 - 10.1016/j.febslet.2011.02.035

DO - 10.1016/j.febslet.2011.02.035

M3 - Article

C2 - 21376045

SN - 0014-5793

VL - 585

SP - 1025

EP - 1030

JO - FEBS letters

JF - FEBS letters

IS - 7

ER -

Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection

Abstract

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