Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection
RT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with Listeria monocytogenes. The pMØ were infected in vitro with L. monocytogenes or vehicle for 4 h. Mice were injected...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/btn-2023-0063 |
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| author | Lethicia Souza Tavares Roberta Lane Oliveira-Silva Marcelo Tigre Moura Jéssica Barboza da Silva Ana Maria Benko-Iseppon José Vitor Lima-Filho |
| author_facet | Lethicia Souza Tavares Roberta Lane Oliveira-Silva Marcelo Tigre Moura Jéssica Barboza da Silva Ana Maria Benko-Iseppon José Vitor Lima-Filho |
| author_sort | Lethicia Souza Tavares |
| collection | DOAJ |
| description | RT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with Listeria monocytogenes. The pMØ were infected in vitro with L. monocytogenes or vehicle for 4 h. Mice were injected with L. monocytogenes (or vehicle) and euthanized 24 h post-injection. The RGs came from a multispecies primer set, from the literature or designed here. The RG ranking relied on GeNorm, NormFinder, BestKeeper, Delta-CT and RefFinder. B2m-H3f3a-Ppia were the most stable RGs for pMØ, albeit RG indexes fine-tuned estimations of cytokine relative expression. Actβ-Ubc-Ppia were the best RGs for spleen but modestly impacted the cytokine relative expression. Hence, mouse models of L. monocytogenes require context-specific RGs for RT-qPCR, thus reinforcing its paramount contribution to accurate gene expression profiling. |
| format | Article |
| id | doaj-art-16ad6e5ff5af4ac7acd47fa90c6f761a |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-16ad6e5ff5af4ac7acd47fa90c6f761a2025-08-20T02:25:34ZengTaylor & Francis GroupBioTechniques0736-62051940-98182023-12-0110.2144/btn-2023-0063Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infectionLethicia Souza Tavares0Roberta Lane Oliveira-Silva1Marcelo Tigre Moura2Jéssica Barboza da Silva3Ana Maria Benko-Iseppon4José Vitor Lima-Filho51Departamento de Biologia, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil2Departamento de Genética, Universidade Federal de Pernambuco, Recife, PE, Brasil3Departamento de Biologia Celular e Molecular, Centro de Biotecnologia, Campus I, Universidade Federal da Paraíba, João Pessoa, PB, Brasil2Departamento de Genética, Universidade Federal de Pernambuco, Recife, PE, Brasil2Departamento de Genética, Universidade Federal de Pernambuco, Recife, PE, Brasil1Departamento de Biologia, Universidade Federal Rural de Pernambuco, Recife, PE, BrasilRT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with Listeria monocytogenes. The pMØ were infected in vitro with L. monocytogenes or vehicle for 4 h. Mice were injected with L. monocytogenes (or vehicle) and euthanized 24 h post-injection. The RGs came from a multispecies primer set, from the literature or designed here. The RG ranking relied on GeNorm, NormFinder, BestKeeper, Delta-CT and RefFinder. B2m-H3f3a-Ppia were the most stable RGs for pMØ, albeit RG indexes fine-tuned estimations of cytokine relative expression. Actβ-Ubc-Ppia were the best RGs for spleen but modestly impacted the cytokine relative expression. Hence, mouse models of L. monocytogenes require context-specific RGs for RT-qPCR, thus reinforcing its paramount contribution to accurate gene expression profiling.https://www.future-science.com/doi/10.2144/btn-2023-0063cytokinehousekeeping genesimmune responseinternal control geneslisteriosismacrophages |
| spellingShingle | Lethicia Souza Tavares Roberta Lane Oliveira-Silva Marcelo Tigre Moura Jéssica Barboza da Silva Ana Maria Benko-Iseppon José Vitor Lima-Filho Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection BioTechniques cytokine housekeeping genes immune response internal control genes listeriosis macrophages |
| title | Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection |
| title_full | Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection |
| title_fullStr | Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection |
| title_full_unstemmed | Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection |
| title_short | Reference genes for gene expression profiling in mouse models of Listeria monocytogenes infection |
| title_sort | reference genes for gene expression profiling in mouse models of listeria monocytogenes infection |
| topic | cytokine housekeeping genes immune response internal control genes listeriosis macrophages |
| url | https://www.future-science.com/doi/10.2144/btn-2023-0063 |
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