Luminescent and fluorescent triple reporter plasmid constructs for Wnt, Hedgehog and Notch pathway Article Swipe
Julia Maier
,
Salma Elmenofi
,
Alexander Taschauer
,
Martina Anton
,
Haider Sami
,
Manfred Ogris
·
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pone.0226570
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pone.0226570
Tracking the activity of signalling pathways is a fundamental method for basic science, as well as in cancer- and pharmaceutical research. The developmental pathways Wnt, Hedgehog and Notch are frequently deregulated in cancers and represent a valuable target for the discovery of novel anticancer compounds. Here we present reporter systems for tracking activity of these pathways by using specific promoter elements driving the expression of either sensitive luciferases or fluorescent proteins. A high level of sensitivity was obtained using the luciferase reporter genes for firefly (FLuc), secreted Gaussia (GLuc) and synthetic NanoLuc (NLuc). As fluorescent reporter proteins, mTurqouise2, tdTomato and iRFP720 were chosen. Specificity of pathway activity was validated by co-transfection with pathway activating genes, showing significant response to induction. In addition, multi-gene plasmids were cloned, allowing the detection of all three pathways by one vector. By using the multi-gene vector 3P-Luc (wnt-NLuc, hedgehog-FLuc, Notch-GLuc), we could unambiguously demonstrate the crosstalk between pathways, while excluding cross reactivity of luciferase substrates. First studies with synthetic compounds confirmed the applicability of the system for future drug screening approaches.
Related Topics
Concepts
Luciferase
Wnt signaling pathway
Hedgehog
Reporter gene
Hedgehog signaling pathway
Biology
Bioreporter
Cell biology
Luciferases
Plasmid
Molecular biology
Transfection
Signal transduction
Gene
Chemistry
Genetics
Gene expression
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1371/journal.pone.0226570
- https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226570&type=printable
- OA Status
- gold
- Cited By
- 12
- References
- 65
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W2994947088
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W2994947088Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1371/journal.pone.0226570Digital Object Identifier
- Title
-
Luminescent and fluorescent triple reporter plasmid constructs for Wnt, Hedgehog and Notch pathwayWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2019Year of publication
- Publication date
-
2019-12-20Full publication date if available
- Authors
-
Julia Maier, Salma Elmenofi, Alexander Taschauer, Martina Anton, Haider Sami, Manfred OgrisList of authors in order
- Landing page
-
https://doi.org/10.1371/journal.pone.0226570Publisher landing page
- PDF URL
-
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226570&type=printableDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
-
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226570&type=printableDirect OA link when available
- Concepts
-
Luciferase, Wnt signaling pathway, Hedgehog, Reporter gene, Hedgehog signaling pathway, Biology, Bioreporter, Cell biology, Luciferases, Plasmid, Molecular biology, Transfection, Signal transduction, Gene, Chemistry, Genetics, Gene expressionTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
12Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 1, 2024: 5, 2023: 1, 2022: 1, 2021: 3Per-year citation counts (last 5 years)
- References (count)
-
65Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.addition, | 121 |
| abstract_inverted_index.compounds | 164 |
| abstract_inverted_index.confirmed | 165 |
| abstract_inverted_index.crosstalk | 150 |
| abstract_inverted_index.detection | 128 |
| abstract_inverted_index.discovery | 40 |
| abstract_inverted_index.excluding | 154 |
| abstract_inverted_index.pathways, | 152 |
| abstract_inverted_index.proteins, | 96 |
| abstract_inverted_index.proteins. | 70 |
| abstract_inverted_index.represent | 34 |
| abstract_inverted_index.research. | 20 |
| abstract_inverted_index.screening | 174 |
| abstract_inverted_index.sensitive | 66 |
| abstract_inverted_index.synthetic | 90, 163 |
| abstract_inverted_index.validated | 108 |
| abstract_inverted_index.(wnt-NLuc, | 142 |
| abstract_inverted_index.activating | 113 |
| abstract_inverted_index.anticancer | 43 |
| abstract_inverted_index.compounds. | 44 |
| abstract_inverted_index.expression | 63 |
| abstract_inverted_index.frequently | 29 |
| abstract_inverted_index.induction. | 119 |
| abstract_inverted_index.luciferase | 80, 158 |
| abstract_inverted_index.multi-gene | 122, 139 |
| abstract_inverted_index.reactivity | 156 |
| abstract_inverted_index.signalling | 4 |
| abstract_inverted_index.Specificity | 103 |
| abstract_inverted_index.approaches. | 175 |
| abstract_inverted_index.demonstrate | 148 |
| abstract_inverted_index.deregulated | 30 |
| abstract_inverted_index.fluorescent | 69, 94 |
| abstract_inverted_index.fundamental | 8 |
| abstract_inverted_index.luciferases | 67 |
| abstract_inverted_index.sensitivity | 75 |
| abstract_inverted_index.significant | 116 |
| abstract_inverted_index.substrates. | 159 |
| abstract_inverted_index.Notch-GLuc), | 144 |
| abstract_inverted_index.mTurqouise2, | 97 |
| abstract_inverted_index.applicability | 167 |
| abstract_inverted_index.developmental | 22 |
| abstract_inverted_index.unambiguously | 147 |
| abstract_inverted_index.hedgehog-FLuc, | 143 |
| abstract_inverted_index.pharmaceutical | 19 |
| abstract_inverted_index.co-transfection | 110 |
| cited_by_percentile_year.max | 98 |
| cited_by_percentile_year.min | 89 |
| corresponding_author_ids | https://openalex.org/A5014456129, https://openalex.org/A5063629000, https://openalex.org/A5024639472, https://openalex.org/A5030915677, https://openalex.org/A5002541021, https://openalex.org/A5083497801 |
| countries_distinct_count | 2 |
| institutions_distinct_count | 6 |
| corresponding_institution_ids | https://openalex.org/I129774422, https://openalex.org/I2802619606, https://openalex.org/I62916508 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/3 |
| sustainable_development_goals[0].score | 0.7400000095367432 |
| sustainable_development_goals[0].display_name | Good health and well-being |
| citation_normalized_percentile.value | 0.61266093 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |