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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...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Library Publishing Media
2010
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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. |
format | Text |
id | pubmed-3043560 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Library Publishing Media |
record_format | MEDLINE/PubMed |
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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