Thread by @SoothSpider

𝑻𝒓𝒂𝒏𝒔-𝒇𝒆𝒄-𝒕𝒊𝒐𝒏
What is a transfection?

"Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods."

Note: viral methods are called "transduction".

Sauce: promega.ca/resources/guid…

Some vehicle is needed because DNA/RNA have the same charge as a cell membrane, making them repulse each other.

Early ideas, like using a cationic polymer (polymers that are positively charged) worked but have short-comings when it comes to transfecting live whole animals (like humans).

Instead Cationic Lipids/Liposomes were developed/designed to overcome those problems.

It is still not clear how exactly a DNA payload crosses the nuclear envelope, but they mostly understand how things get into the cytoplasm.

Transaction Reagents have been created that work w/ a wide variety of eukaryotic cell lines. (Meaning they work w/ cells that are not bacteria – for ex – & are traditionally more resistant to taking in foreign material.)

Side thought: Why do bacteria readily take in material?

Nonliposomal methods have also been created.

Side Note: Cullis? Pieter Cullis? 🤔

The original transfection methods were Physical Methods including microinjection, electroporation and particle bombardment (the "pew pew" method).

It's interesting how in the shiny product brochure they reiterate several times how various transfection reagents have "low toxicity", but then emphasize that something has to be done about the toxicity of the reagent. 🤔🤔🤔

Just a side note in most biological measurements, you NORMALLY need a "reference" measurement. Bc many things are changing, how does the researcher know if the expression of what they're looking for is going up or down when everything is going up?

Yes, looking at you PCR tests.

So in the end, transfections are not new. They are a product that has been used in experimentation for a long time. They were classically used for research needing gene transfers, but have also been used for a variety of different things. Most bench biologists would know this.

@threadreaderapp unroll