Can you transfect neurons?
Transfection is the process of introducing nucleic acids into cells. The term transfection is commonly used for non-viral gene delivery (1, 2). Electroporation is generally used with freshly isolated neurons or neuronal cell lines in suspension, and requires specialized equipment which is relatively expensive.
What is genetic transfection?
Broadly defined, transfection is the process of artificially introducing nucleic acids (DNA or RNA) into cells, utilizing means other than viral infection.
What is chemical transfection?
Transfection is the introduction of any nucleic acid molecule by non-viral means into cultured eukaryotic cells. In the past, it was often used only in reference to DNA, but that has changed as applications such as RNAi and more recently, CRISPR have been developed.
What is the difference between Nucleofection and electroporation?
With its superior transfection performance, Nucleofection offers various advantages over traditional electroporation methods: High transfection efficiencies of up to 90% for plasmid DNA and 99% for oligonucleotides, like siRNA. Excellent preservation of the physiological status and viability of transfected cells.
What is a transfection used for?
The main purpose of transfection is to study the function of genes or gene products, by enhancing or inhibiting specific gene expression in cells, and to produce recombinant proteins in mammalian cells [3].
What is the process of transfection?
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Transfection of animal cells typically involves opening transient pores or “holes” in the cell membrane to allow the uptake of material.
What are the types of transfection?
Some of the commonly used transfection techniques include calcium phosphate precipitation, lipofection, electroporation, and viral delivery.
Can a lipid be used to transfect primary neurons?
However, the same lipids, when used to transfect postmitotic neurons, tend to give poorer results (typically 1–5%), although maximum values of up to 30% have been reported for primary neurons ( Dalby et al., 2004 ). These comparatively low transfection efficiencies]
What is the efficiency of transfection in neurons?
In postmitotic cells such as neurons, entry into the nucleus is more difficult and the expression rate consequently reduced. Therefore, the transfection efficiency generally lies between 1 and 5% and, even after optimization, rarely reaches 30% ( Goetze et al., 2004 ).
Which is the best cell specific transfection protocol?
Cell-Specific Transfection Protocols NOTE: Invitrogen Lipofectamine Transfection Reagent Protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. This is often the best place to start, especially in a new cell line.
How is plasmid DNA used to transfect neurons?
The neuron is subsequently electroporated with a high resistance patch pipette containing the plasmid DNA. Following the electroporation, transfected neurons can be imaged and/or targeted for whole-cell patch-clamp recordings.