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Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes

Transgenic technologies can provide important animal models for studying drug-metabolizing enzymes. Our overall aim was to generate versatile cell and animal systems that exhibited varying levels of cytochrome P450 oxidoreductase (POR) activity, more accurately modelling the human population for pha...

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Detalles Bibliográficos
Autores principales: Mackenzie, Neil CW, Lillico, Simon G, Brown, Ken, Wolf, Charles R, Whitelaw, Christopher BA
Formato: Texto
Lenguaje:English
Publicado: Library Publishing Media 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3043560/
https://www.ncbi.nlm.nih.gov/pubmed/21350684
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author Mackenzie, Neil CW
Lillico, Simon G
Brown, Ken
Wolf, Charles R
Whitelaw, Christopher BA
author_facet Mackenzie, Neil CW
Lillico, Simon G
Brown, Ken
Wolf, Charles R
Whitelaw, Christopher BA
author_sort Mackenzie, Neil CW
collection PubMed
description Transgenic technologies can provide important animal models for studying drug-metabolizing enzymes. Our overall aim was to generate versatile cell and animal systems that exhibited varying levels of cytochrome P450 oxidoreductase (POR) activity, more accurately modelling the human population for pharmacological and toxicology studies. Towards this goal we evaluated RNA-interference constructs designed for use in vitro and in vivo for reducing POR activity in hepatocytes. This study clearly demonstrates that both POR protein level and reductase activity can be significantly knocked down in Hepa-1 cells in vitro, while highlighting the difficulty in predicting knockdown efficiency in transgenic animals. The high levels of embryonic lethality observed, and inability to produce multi-copy transgenic animals indicates that high levels of shRNA expression may be detrimental to embryonic development.
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spelling pubmed-30435602011-02-24 Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes Mackenzie, Neil CW Lillico, Simon G Brown, Ken Wolf, Charles R Whitelaw, Christopher BA J RNAi Gene Silencing Research Report Transgenic technologies can provide important animal models for studying drug-metabolizing enzymes. Our overall aim was to generate versatile cell and animal systems that exhibited varying levels of cytochrome P450 oxidoreductase (POR) activity, more accurately modelling the human population for pharmacological and toxicology studies. Towards this goal we evaluated RNA-interference constructs designed for use in vitro and in vivo for reducing POR activity in hepatocytes. This study clearly demonstrates that both POR protein level and reductase activity can be significantly knocked down in Hepa-1 cells in vitro, while highlighting the difficulty in predicting knockdown efficiency in transgenic animals. The high levels of embryonic lethality observed, and inability to produce multi-copy transgenic animals indicates that high levels of shRNA expression may be detrimental to embryonic development. Library Publishing Media 2010-11-26 /pmc/articles/PMC3043560/ /pubmed/21350684 Text en ©The Authors http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an open access article, published under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/). This license permits non-commercial use, distribution and reproduction of the article, provided the original work is appropriately acknowledged with correct citation details.
spellingShingle Research Report
Mackenzie, Neil CW
Lillico, Simon G
Brown, Ken
Wolf, Charles R
Whitelaw, Christopher BA
Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title_full Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title_fullStr Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title_full_unstemmed Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title_short Evaluation of RNA-knockdown strategies for modulation of cytochrome P450 reductase activity in mouse hepatocytes
title_sort evaluation of rna-knockdown strategies for modulation of cytochrome p450 reductase activity in mouse hepatocytes
topic Research Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3043560/
https://www.ncbi.nlm.nih.gov/pubmed/21350684
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