Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia
Summary: In this protocol, we describe the small interfering RNA (siRNA)-mediated gene knockdown in primary mouse microglia, providing an approach to investigate functions such as phagocytosis and chemotaxis. The approach includes siRNA design, establishment of mixed glial cultures, microglia isolat...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724000327 |
| _version_ | 1797317154409807872 |
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| author | Yuma Kato Sho Takatori Aika Akahori Hayato Etani Yung Ning Chu Taisuke Tomita |
| author_facet | Yuma Kato Sho Takatori Aika Akahori Hayato Etani Yung Ning Chu Taisuke Tomita |
| author_sort | Yuma Kato |
| collection | DOAJ |
| description | Summary: In this protocol, we describe the small interfering RNA (siRNA)-mediated gene knockdown in primary mouse microglia, providing an approach to investigate functions such as phagocytosis and chemotaxis. The approach includes siRNA design, establishment of mixed glial cultures, microglia isolation, and siRNA transfection. Validation of knockdown efficacy employs quantitative immunoblot analysis. This technique empowers the investigation of specific molecular and cellular functions within the intricate microenvironment of the brain, comprising diverse cell types.For complete details on the use and execution of this protocol, please refer to Iguchi et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T03:29:47Z |
| format | Article |
| id | doaj.art-e1d3625647c3498b81bfb9d687a80654 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T03:29:47Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-e1d3625647c3498b81bfb9d687a806542024-02-11T05:12:09ZengElsevierSTAR Protocols2666-16672024-03-0151102867Protocol for gene knockdown using siRNA in primary cultured neonatal murine microgliaYuma Kato0Sho Takatori1Aika Akahori2Hayato Etani3Yung Ning Chu4Taisuke Tomita5Laboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Corresponding authorSummary: In this protocol, we describe the small interfering RNA (siRNA)-mediated gene knockdown in primary mouse microglia, providing an approach to investigate functions such as phagocytosis and chemotaxis. The approach includes siRNA design, establishment of mixed glial cultures, microglia isolation, and siRNA transfection. Validation of knockdown efficacy employs quantitative immunoblot analysis. This technique empowers the investigation of specific molecular and cellular functions within the intricate microenvironment of the brain, comprising diverse cell types.For complete details on the use and execution of this protocol, please refer to Iguchi et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724000327Cell BiologyCell isolationImmunologyMolecular BiologyGene Expression |
| spellingShingle | Yuma Kato Sho Takatori Aika Akahori Hayato Etani Yung Ning Chu Taisuke Tomita Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia STAR Protocols Cell Biology Cell isolation Immunology Molecular Biology Gene Expression |
| title | Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia |
| title_full | Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia |
| title_fullStr | Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia |
| title_full_unstemmed | Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia |
| title_short | Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia |
| title_sort | protocol for gene knockdown using sirna in primary cultured neonatal murine microglia |
| topic | Cell Biology Cell isolation Immunology Molecular Biology Gene Expression |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000327 |
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