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A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues
BACKGROUND: Analogues of vitamin D3 are extensively used in the treatment of various illnesses, such as osteoporosis, inflammatory skin diseases, and cancer. Functional testing of new vitamin D3 analogues and formulations for improved systemic and topical administration is supported by sensitive scr...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3083354/ https://www.ncbi.nlm.nih.gov/pubmed/21473770 http://dx.doi.org/10.1186/1472-6750-11-33 |
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author | Staunstrup, Nicklas H Sharma, Nynne Bak, Rasmus O Svensson, Lars Petersen, Thomas K Aarenstrup, Lene Kristiansen, Karsten Bolund, Lars Mikkelsen, Jacob Giehm |
author_facet | Staunstrup, Nicklas H Sharma, Nynne Bak, Rasmus O Svensson, Lars Petersen, Thomas K Aarenstrup, Lene Kristiansen, Karsten Bolund, Lars Mikkelsen, Jacob Giehm |
author_sort | Staunstrup, Nicklas H |
collection | PubMed |
description | BACKGROUND: Analogues of vitamin D3 are extensively used in the treatment of various illnesses, such as osteoporosis, inflammatory skin diseases, and cancer. Functional testing of new vitamin D3 analogues and formulations for improved systemic and topical administration is supported by sensitive screening methods that allow a comparative evaluation of drug properties. As a new tool in functional screening of vitamin D3 analogues, we describe a genomically integratable sensor for sensitive drug detection. This system facilitates assessment of the pharmacokinetic and pharmadynamic properties of vitamin D3 analogues. The tri-cistronic genetic sensor encodes a drug-sensoring protein, a reporter protein expressed from an activated sensor-responsive promoter, and a resistance marker. RESULTS: The three expression cassettes, inserted in a head-to-tail orientation in a Sleeping Beauty DNA transposon vector, are efficiently inserted as a single genetic entity into the genome of cells of interest in a reaction catalyzed by the hyperactive SB100X transposase. The applicability of the sensor for screening purposes is demonstrated by the functional comparison of potent synthetic analogues of vitamin D3 designed for the treatment of psoriasis and cancer. In clones of human keratinocytes carrying from a single to numerous insertions of the vitamin D3 sensor, a sensitive sensor read-out is detected upon exposure to even low concentrations of vitamin D3 analogues. In comparative studies, the sensor unveils superior potency of new candidate drugs in comparison with analogues that are currently in clinical use. CONCLUSIONS: Our findings demonstrate the use of the genetic sensor as a tool in first-line evaluation of new vitamin D3 analogues and pave the way for new types of drug delivery studies in sensor-transgenic animals. |
format | Text |
id | pubmed-3083354 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-30833542011-04-28 A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues Staunstrup, Nicklas H Sharma, Nynne Bak, Rasmus O Svensson, Lars Petersen, Thomas K Aarenstrup, Lene Kristiansen, Karsten Bolund, Lars Mikkelsen, Jacob Giehm BMC Biotechnol Research Article BACKGROUND: Analogues of vitamin D3 are extensively used in the treatment of various illnesses, such as osteoporosis, inflammatory skin diseases, and cancer. Functional testing of new vitamin D3 analogues and formulations for improved systemic and topical administration is supported by sensitive screening methods that allow a comparative evaluation of drug properties. As a new tool in functional screening of vitamin D3 analogues, we describe a genomically integratable sensor for sensitive drug detection. This system facilitates assessment of the pharmacokinetic and pharmadynamic properties of vitamin D3 analogues. The tri-cistronic genetic sensor encodes a drug-sensoring protein, a reporter protein expressed from an activated sensor-responsive promoter, and a resistance marker. RESULTS: The three expression cassettes, inserted in a head-to-tail orientation in a Sleeping Beauty DNA transposon vector, are efficiently inserted as a single genetic entity into the genome of cells of interest in a reaction catalyzed by the hyperactive SB100X transposase. The applicability of the sensor for screening purposes is demonstrated by the functional comparison of potent synthetic analogues of vitamin D3 designed for the treatment of psoriasis and cancer. In clones of human keratinocytes carrying from a single to numerous insertions of the vitamin D3 sensor, a sensitive sensor read-out is detected upon exposure to even low concentrations of vitamin D3 analogues. In comparative studies, the sensor unveils superior potency of new candidate drugs in comparison with analogues that are currently in clinical use. CONCLUSIONS: Our findings demonstrate the use of the genetic sensor as a tool in first-line evaluation of new vitamin D3 analogues and pave the way for new types of drug delivery studies in sensor-transgenic animals. BioMed Central 2011-04-07 /pmc/articles/PMC3083354/ /pubmed/21473770 http://dx.doi.org/10.1186/1472-6750-11-33 Text en Copyright ©2011 Staunstrup et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Staunstrup, Nicklas H Sharma, Nynne Bak, Rasmus O Svensson, Lars Petersen, Thomas K Aarenstrup, Lene Kristiansen, Karsten Bolund, Lars Mikkelsen, Jacob Giehm A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title | A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title_full | A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title_fullStr | A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title_full_unstemmed | A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title_short | A Sleeping Beauty DNA transposon-based genetic sensor for functional screening of vitamin D3 analogues |
title_sort | sleeping beauty dna transposon-based genetic sensor for functional screening of vitamin d3 analogues |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3083354/ https://www.ncbi.nlm.nih.gov/pubmed/21473770 http://dx.doi.org/10.1186/1472-6750-11-33 |
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