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wanson t1_iy2uc5j wrote

Reply to comment by Aescwicca in How exactly does CRISPR-CAS9 insert new genes? by AutomaticAd1918

It’s just an oligo. We use them as primers all the time in PCR. You just decide what bases you want and order it from a company. Small 20 base pair primers cost about $3.

They’re made by a chemical process and shipped dried (lyophilized). We just dissolve them in water or buffer.

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HotDadBod1255 t1_iy36fb7 wrote

Oligos used for CRISPR-Cas9 are extremely expensive. They're usually larger (60-120mer) and have to be much more pure to avoid off target editing.

They're currently one of the biggest bottlenecks in the gene therapy field since we can't make enough of it fast enough and with high enough purity.

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wanson t1_iy3hkaj wrote

They can be. It depends on what you're doing. For me, I am just correcting SNPs in cell lines so 50-60 bp is enough and they're only a couple hundred dollars or so.

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HotDadBod1255 t1_iy3l7ih wrote

Right, totally different needs here. I use them for in-vivo gene editing, so purity and precision are paramount.

Like most things out there, when you need super high purity and quality, it's gonna cost you a lot. In this case oligos are already a pain to make so it's ridiculously expensive.

To give you an idea, around 2000mg of the oligo I use as guide RNA will cost in the $2M range.

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