I am a Master's student now and I am currently working on a project related to mRNA loading lipid nanoparticles and I want to check the translation ability of mRNA on HEK293T cell lines (I will perform the Western-Blot too). That's why I need to fuse mRNA to Nano-luciferase protein for Nano-Luciferase assay. The Biotech company where I contacted that also recommended me 2 kinds of linker (GGGGS and GGSGGT) but in the most articles I read related to my intended mRNA, no one mentioned which kind of linker they used. Because my previous background is not biology, these days I am really in panic.
I also attached my intended mRNA sequence below. Thank you in advance for your advice. It will be so much meaningful to me.
https://www.ncbi.nlm.nih.gov/nuccore/NM_007162.3?report=fasta&to=2354
When designing a fusion protein, the linker sequence plays a crucial role in maintaining the functionality and stability of both the individual protein domains and the fusion construct as a whole. For the fusion of NanoLuciferase (NanoLuc) with an mRNA, you'll want a linker that provides flexibility, minimizes steric hindrance, and ideally allows for efficient translation of the mRNA.
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