I'm going to use cells infected with HIV-based translentiviral recombinant vectors containing the human wt- or F508del-CFTR cDNAs. Does anyone know which biosafety level is required to use this kind of cells? Could these cells be use safely for patch clamp experiments?
Hi Maria,
I can only comment on UK regulations:
In our institute the biosafety level of lentiviral work depends on the transgenes encoded within the virus, ie expression of oncogenes or knock-down of tumour suppressor genes are regarded as class 2, and all other transgenes are class 1. All class 2 work is performed at biosafety level 2. This includes all virus production, concentration and infection that involves a class 2 virus. After infection we have 'local rules' at our institue based on data I generated that say after 4 washes the infected cells become class 1 (unless there is a specific reason to keep them in class 2).
It will all depend on where you are and what the rules are in your institute. As far as safety goes, the risk of generating replication competent viruses with a 3rd gen system is virtually non-existent (and this particular system you're working with is touted by OpenBio for its safety), so I wouldn't let that worry you.
Good luck.
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