Effective Knockdown of Gene Expression in Primary Microglia With siRNA and Magnetic Nanoparticles Without Cell Death or Inflammation.
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Authors
Carrillo-Jimenez, Alejandro
Puigdellívol, Mar
Vilalta, Anna
Venero, Jose Luis
Brown, Guy Charles
StGeorge-Hyslop, Peter
Publication Date
2018Journal Title
Front Cell Neurosci
ISSN
1662-5102
Publisher
Frontiers Media SA
Volume
12
Pages
313
Language
eng
Type
Article
Physical Medium
Electronic-eCollection
Metadata
Show full item recordCitation
Carrillo-Jimenez, A., Puigdellívol, M., Vilalta, A., Venero, J. L., Brown, G. C., StGeorge-Hyslop, P., & Burguillos, M. A. (2018). Effective Knockdown of Gene Expression in Primary Microglia With siRNA and Magnetic Nanoparticles Without Cell Death or Inflammation.. Front Cell Neurosci, 12 313. https://doi.org/10.3389/fncel.2018.00313
Abstract
Microglia, the resident immune cells of the brain, have multiple functions in physiological and pathological conditions, including Alzheimer's disease (AD). The use of primary microglial cell cultures has proved to be a valuable tool to study microglial biology under various conditions. However, more advanced transfection methodologies for primary cultured microglia are still needed, as current methodologies provide low transfection efficiency and induce cell death and/or inflammatory activation of the microglia. Here, we describe an easy, and effective method based on the Glial-Mag method (OZ Biosciences) using magnetic nanoparticles and a magnet to successfully transfect primary microglia cells with different small interfering RNAs (siRNAs). This method does not require specialist facilities or specific training and does not induce cell toxicity or inflammatory activation. We demonstrate that this protocol successfully decreases the expression of two key genes associated with AD, the triggering receptor expressed in myeloid cells 2 (TREM2) and CD33, in primary microglia cell cultures.
Sponsorship
European Commission Horizon 2020 (H2020) Research Infrastructures (RI) (115976)
Identifiers
External DOI: https://doi.org/10.3389/fncel.2018.00313
This record's URL: https://www.repository.cam.ac.uk/handle/1810/285810
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