Dear Hanna!

What vector do You want to use? What promoter?

Hello

I'm sorry, this is not my field, but I am very interested in helping researchers in the field of genetics.

Can I help with this plan with just a few simple searches? like this

https://openwetware.org/wiki/Griffin:Puromycin_Selection

Article A transgenic mouse embryonic stem cell line for puromycin se...

Thanks for trying to help me! The promoter we are planning to use to drive puromycin expression is EFS, which is derived from EF1a by removing an intron. I know that EF1a works pretty OK in neurons, so it may be strong enough to protect the cell during selection. I just need to test it together with the puromycin kill curve. I have also found now three papers, where they have performed puromycin selection in primary neurons:

1) 10.1016/j.bbamcr.2009.07.001

2) 10.1371/journal.pone.0091744.

3) doi.org/10.1016/j.isci.2022.105695

The paper that Seyed Hossein Ahmadi recommended used mESC line. In most cases, puromycin selection has been done in cell lines or some type of proliferating cells such as NPC, mESC, iPSC before differentiating them into neurons. But given that I have already found three papers that have performed puro selection in cultured primary neurons gives me hope that this will be feasible.

Thanks again!

Which transgenesis system You want to use?

Adam Figarski Planning of running a CRISPR screen in primary neurons and using puromycin selection for transduced neurons.

As Soyed mentioned the paper it should work, I just wonder if You need mature neurons to be transduced ( so ----> genetically modified) or neuroblasts are also fine an You can differentiate them after putting library? in.