Hi everyone,

I’m trying to design a plasmid that, in combination with a viral vector, might prevent the involution of the thymus in humans. I’m going off this paper where they gave some mice a copy of the foxN1 gene, but with a keratin-14 promoter instead of whatever promoter foxn1 has in nature: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3228493/

Only problem is, it looks like they modified the dna of some embryos, whereas I want my mod to work on an already-developed organism. I would just put the foxN1-k14 sequence in a lentiviral backbone and call it a day, but HIV-1 can only carry 8-12 kilobases, and human foxN1 has like 15kb.

I can’t seem to find any viral vectors that can hold this much code, but it looks like I might be able to split up my payload into two separate viral vectors, and have them recombine once they’re in the cell, using an intein.

I read about an intein that is commonly used in mammalian cells called Ssp DnaE, but I can’t find any actual code for it that I can copy and paste into benchling. Also, I have no idea how it needs to be inserted. Once I have it, do I just plop it at the end of each half of the foxN1-k14 sequence and hope for the best? Or is there a more sophisticated implementation?

Thanks