HSV-1 measurement of cell-cell fusion

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  • Last Post 08 May 2017
Jelste posted this 08 May 2017

Hello,

Our group is interested in a luciferase reporter system to measure cell-cell fusion between effector cells (CHO-K1) transfected with HSV-1 glycoproteins and target cells (HeLa). We are testing a compound that blocks viral entry/fusion and we simply want to measure the amount of fusion occurring in relation to a positive and negative control. For this application, would the NanoBit inducible control pair be an acceptable choice?

Thank you,

Jim

alandrem posted this 08 May 2017

Hello Jim,

Thank you for your interest in using NanoBiT for your cell fusion assay. I do think you could use NanoBiT for this application, but the high affinity system might be your best bet. For example, you could engineer your HeLa cells to express the LgBiT subunit and you could engineer your CHO-K1 cells to express the HiBiT (high affinity peptide). If a cell fusion even occurred the HiBiT and LgBiT would spontaneously complement in order to give the active enzyme. However, if your compounds blocked fusion you would not get the enzyme and so would not get the bioluminescence. We currently have other researchers evaluating the high affinity system for cell fusion and/or viral infectivity assays. There are certainly some things to consider (e.g. expression levels), but we think this should work. Let me know if you are interested in giving it a try.

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