ArticleJournal of neuroscience methods2020
A highly efficient method for single-cell electroporation in mouse organotypic hippocampal slice culture.
Article in Journal of neuroscience methods, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- CRISPR/Cas9 systems: Delivery technologies and biomedical applications.Asian journal of pharmaceutical sciences · 2023Review
- Regulation of Presynaptic Release Machinery by Cell Adhesion Molecules.Advances in neurobiology · 2023Article
- Deep Learning-Assisted Automated Single Cell Electroporation Platform for Effective Genetic Manipulation of Hard-to-Transfect Cells.Small (Weinheim an der Bergstrasse, Germany) · 2022Article
- Neuroligin-3: A Circuit-Specific Synapse Organizer That Shapes Normal Function and Autism Spectrum Disorder-Associated Dysfunction.Frontiers in molecular neuroscience · 2021Review
- Specific Neuroligin3-αNeurexin1 signaling regulates GABAergic synaptic function in mouse hippocampus.eLife · 2020Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
backgroundExogenous gene introduction by transfection is one of the most important approaches for understanding the function of specific genes at the cellular level. Electroporation has a long-standing history as a versatile gene delivery technique in vitro and in vivo. However, it has been underutilized in vitro because of technical difficulty and insufficient transfection efficiency. NEW
methodWe have developed an electroporation technique that combines the use of large glass electrodes, tetrodotoxin-containing artificial cerebrospinal fluid and mild electrical pulses. Here, we describe the technique and compare it with existing methods.
resultsOur method achieves a high transfection efficiency (∼80 %) in both excitatory and inhibitory neurons with no detectable side effects on their function. We demonstrate this method is capable of transferring at least three different genes into a single neuron. In addition, we demonstrate the ability to transfect different genes into neighboring cells. COMPARISON WITH EXISTING
methodsThe majority of existing methods use fine-tipped glass electrodes (i.e. > 10 MΩ) and apply high voltage (10 V) pulses with high frequency (100 Hz) for 1 s. These parameters contribute to practical difficulties thus lowering the transfection efficiency. Our unique method minimizes electrode clogging and therefore procedure duration, increasing transfection efficiency and cellular viability.
conclusionsOur modifications, relative to current methods, optimize electroporation efficiency and cell survival. Our approach offers distinct research strategies not only in elucidating cell-autonomous functions of genes but also for assessing genes contributing to intercellular functions, such as trans-synaptic interactions.
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