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^*

^*Corresponding author for this work

University of Amsterdam

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

25 Citations (Scopus)

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
Number of pages	6
Journal	FEBS Letters. Journal for the Rapid Publication of Short Reports in Biochemistry .....
Volume	585
Issue number	7
DOIs	https://doi.org/10.1016/j.febslet.2011.02.035
Publication status	Published - 6 Apr 2011

Bibliographical note

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EMC MM-03-47-02-A
EMC MM-04-20-01
EMC MM-04-20-02-A
EMC MM-04-47-07

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10.1016/j.febslet.2011.02.035

http://hdl.handle.net/1765/25537

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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. Journal for the Rapid Publication of Short Reports in Biochemistry ....., 585(7), 1025-1030. https://doi.org/10.1016/j.febslet.2011.02.035

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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\}",

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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. Journal for the Rapid Publication of Short Reports in Biochemistry ....., vol. 585, no. 7, pp. 1025-1030. https://doi.org/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. / Pan, Qiuwei; de Ruiter, Petra E; von Eije, Karin J et al.
In: FEBS Letters. Journal for the Rapid Publication of Short Reports in Biochemistry ....., Vol. 585, No. 7, 06.04.2011, p. 1025-1030.

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

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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

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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.

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Pan Q, de Ruiter PE, von Eije KJ, Smits R, Kwekkeboom J, Tilanus HW et al. 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. Journal for the Rapid Publication of Short Reports in Biochemistry ..... 2011 Apr 6;585(7):1025-1030. 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

Abstract

Bibliographical note

Research programs

UN SDGs

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