Transfection Techniques for Neuronal Cells: Table 1.
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Abstract Macrophages populate every organ during homeostasis and disease, displaying features of tissue imprinting and heterogeneous activation. The disjointed picture of macrophage biology that emerged from these observations are a barrier for integration across models or with in vitro macrophage activation paradigms. We set out to contextualize macrophage heterogeneity across mouse tissues and inflammatory conditions, specifically aiming to define a common framework of macrophage activation. We built a predictive model with which we mapped the activation of macrophages across 12 tissues and 25 biological conditions, finding a striking commonality and finite number of transcriptional profiles, which we modelled as defined stages along four conserved activation paths. We verified this model with adoptive cell transfer experiments and identified transient RELMɑ expression as a feature of macrophage tissue engraftment. We propose that this integrative approach of macrophage classification allows the establishment of a common predictive framework of macrophage activation in inflammation and homeostasis. One Sentence Summary We propose an integrative approach of macrophage classification that allows the establishment of a common framework of macrophage activation in inflammation and homeostasis.
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LNPs are much more efficient and less toxic than current methods used to deliver siRNA to cultured neurons such as electroporation, calcium phosphate, or lipofection.
“These results indicate that LNPs are much more efficient and less toxic than current methods used to deliver siRNA to cultured neurons such as electroporation, calcium phosphate, or lipofection, which usually result in transfection rates of 1–10%.13”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [13]
The Lipofection section reports siRNA delivery into postmitotic neurons at up to 83% efficiency, contrary to the cited sentence’s characterization of siRNA-delivery methods as usually achieving 1–10%. Its low lipofection rates of typically 1–5% are explicitly for plasmid-based vectors, not siRNA. The paper does not compare LNP efficiency or toxicity with these methods. (LLM, conf=0.88)
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Experimental — accuracy not yet measuredCurrent nonviral vectors exhibit transfection efficiencies of 10% or less and are often accompanied by cell damage.
“By contrast, current nonviral vectors exhibit transfection efficiencies of 10% or less and are often accompanied by cell damage.13”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [13]
The Electrical transfection methods section directly contradicts the claim’s 10% ceiling: it reports nucleofection rates averaging 60–80% after optimization, with up to 95% in embryonic rat neurons and better cell survival than conventional electroporation. The paper does report cell damage with some nonviral techniques, including biolistics, but does not characterize nonviral methods as uniformly low-efficiency and damaging. Page numbers were not available in our copy. (LLM, conf=0.90)
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Experimental — accuracy not yet measuredAAV has limitations in some gene therapy studies, such as toxicity and unnecessary immunoreaction, especially in neuroscience research.
“AAV also has been shown to have limitations in a few gene therapy studies, such as its toxicity and unnecessary immunoreaction, especially in neuroscience research [66,67].”
From Non-Integrating Lentiviral Vectors in Clinical Applications: A Glance Through (2022) · cites this paper as [67]
The Adeno-associated viruses row of Table 1 lists immune or inflammatory responses in vivo as an AAV limitation, supporting that part of the claim. But it rates AAV toxicity as low, and the Virus-based transfection methods section attributes possible cytotoxic effects to contaminating helper viruses rather than AAV itself. The supplied passages have no page numbers. (LLM, conf=0.87)
Experimental — accuracy not yet measuredViral delivery is the most effective available way to transfect neurons.
“Viral delivery is the most effective available way to transfect neurons.13,24”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [13]
The “Virus-based transfection methods” section describes viral delivery as extremely efficient and often the only viable option for efficient delivery to specific cell types in vivo. It does not call viral delivery the most effective method for neurons generally; the “Electrical transfection methods” section instead calls nucleofection the method of choice when high transfection efficiency is essential. (LLM, conf=0.85)
Experimental — accuracy not yet measuredCurrent methods to deliver siRNA to cultured neurons (electroporation, calcium phosphate, or lipofection) usually result in transfection rates of 1-10%.
“which usually result in transfection rates of 1–10%.13”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [13]
The calcium-phosphate table entry gives typical efficiencies of 5–10% in postmitotic neurons, and the Lipofection section gives 1–5% for plasmid-based transfection. Those figures do not establish the claimed usual rate for siRNA delivery: the Lipofection section instead describes efficient siRNA transfer, reaching up to 83% in primary rat hippocampal neurons. (LLM, conf=0.86)
Experimental — accuracy not yet measured
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Cites
Rahim, A. A., Wong, A. M. S., Howe, S. J., Buckley, S. M. K., Acosta-Saltos, A. D., Elston, K. E., Ward, N. J., Philpott, N. J., Cooper, J. D., Anderson, P. N., et al. (2009) · Gene Therapy · doi:10.1038/gt.2008.186 · Cited by 101 · see citing works
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Research works printed but not linked: 3
- Epstein AL (2009) HSV-1-derived amplicon vectors: recent technological improvements and remaining difficulties-a review. Mem Inst Oswaldo Cruz 104:399 -410.
- Fraefel C (2002) Gene delivery using helper virus-free HSV-1 amplicon vectors. Curr Pro- toc Hum Genet Chapter 12:Unit 12.12. Goetze B, Grunewald B, Baldassa S, Kiebler M (2004) Chemically controlled formation of a DNA/calcium phosphate coprecipitate: appli- cation for transfection of mature hippocam- pal neurons. J Neurobiol 60:517-525.
- Zeitelhofer M, Vessey JP, Thomas S, Kiebler M, Dahm R (2009b) Transfection of cultured primary neurons via nucleofection. Curr Pro- toc Neurosci Chapter 4:Unit 4.
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In vitro and ex vivo strategies for intracellular delivery
Stewart, M. P., Sharei, A., Ding, X., Sahay, G., Langer, R., & Jensen, K. F. (2016) · Nature · doi:10.1038/nature19764 · Cited by 885 · see citing works
Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain
Rungta, R. L., Choi, H. B., Lin, P. J., Ko, R. W., Ashby, D., Nair, J., Manoharan, M., Cullis, P. R., & MacVicar, B. A. (2013) · Molecular Therapy—Nucleic Acids · doi:10.1038/mtna.2013.65 · Cited by 165 · see citing works
Hydrophobically Modified siRNAs Silence Huntingtin mRNA in Primary Neurons and Mouse Brain
Alterman, J. F., Hall, L. M., Coles, A. H., Hassler, M. R., Didiot, M.-C., Chase, K., Abraham, J., Sottosanti, E., Johnson, E., Sapp, E., et al. (2015) · Molecular Therapy - Nucleic Acids · doi:10.1038/mtna.2015.38 · Cited by 158 · see citing works
An organotypic brain slice preparation from adult patients with temporal lobe epilepsy
Eugène, E., Cluzeaud, F., Cifuentes-Diaz, C., Fricker, D., Le Duigou, C., Clemenceau, S., Baulac, M., Poncer, J.-C., & Miles, R. (2014) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2014.07.009 · Cited by 101 · see citing works
Non-Integrating Lentiviral Vectors in Clinical Applications: A Glance Through
Gurumoorthy, N., Nordin, F., Tye, G. J., Wan Kamarul Zaman, W. S., & Ng, M. H. (2022) · Biomedicines · doi:10.3390/biomedicines10010107 · Cited by 69 · see citing works
- Authors
- Karra, D., & Dahm, R.
- Affiliations
- Heinrich Heine University Düsseldorf (DE)
- University of Padua (IT)
- Corresponding author
- Dahm, Ralf
- ORCID
- Dahm, Ralf
- Venue
- The Journal of Neuroscience
- Publisher
- Society for Neuroscience
- Published
- 2010-05-05
- Volume
- 30
- Issue
- 18
- Pages
- 6171-6177
- ISSN
- 0270-6474 (ISSN-L)
- 1529-2401
- PMC
- PMC6632737
- OpenAlex
- W2113811929
- Cited by
- 209
- References
- 1
- Citations by year
- 2026: 8 · 2025: 9 · 2024: 18 · 2023: 7 · 2022: 9 · 2021: 11 · 2020: 10 · 2019: 18 · 2018: 15 · 2017: 13 · 2016: 21 · 2015: 20 · 2014: 12 · 2013: 20 · 2012: 10
- FWCI
- 7.37
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- neurons, macrophage heterogeneity, macrophage activation, RELMα, gene expression profiling, inflammation, homeostasis, adoptive cell transfer
- MeSH terms
- Animals, Humans, Neurons, Transduction, Genetic, Transfection, Electroporation, Biolistics
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