It might be cleaner to use an IRES sequence right after the stop codon of SYCP3 so your fluorescent reporter expresses off the same transcript without being conjugated directly to SYCP3. Would be an issue if you were imaging for SYCP3 localization, but you just want flow sorting, right?
I actually tried that with SYCP1 in late 2020/early 2021. The IRES really diminishes the expression of the fluorescent protein and it was undetectable by flow cytometry, even when I used CRISPRa to activate expression of the RNA. See also: https://www.sciencedirect.com/science/article/pii/S1525001600900509
It might be cleaner to use an IRES sequence right after the stop codon of SYCP3 so your fluorescent reporter expresses off the same transcript without being conjugated directly to SYCP3. Would be an issue if you were imaging for SYCP3 localization, but you just want flow sorting, right?
It's a pretty well-known technology and I don't know the literature very well for it, but a bit of searching was able to find me this: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2275123/
I actually tried that with SYCP1 in late 2020/early 2021. The IRES really diminishes the expression of the fluorescent protein and it was undetectable by flow cytometry, even when I used CRISPRa to activate expression of the RNA. See also: https://www.sciencedirect.com/science/article/pii/S1525001600900509
Ah, RIP.