Zeomycin resistance gene in lentiCRISPR v.2

ajanssen3

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Feb 19, 2016, 4:16:17 PM2/19/16

to Genome Engineering using CRISPR/Cas Systems

In lentiCRISPR v.2 plasmid, there is a Zeomycin resistance gene ("bleo," between 12045 and 12416), after an SV40 promoter. Does anyone know why this gene is there and at what point in the process would you use it for selection? Would this be used during leniviral production? Or the creation of a stable cell line?

Any insight is appreciated. Thanks.

Anita

ajanssen3

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Feb 19, 2016, 4:18:58 PM2/19/16

to Genome Engineering using CRISPR/Cas Systems

Sorry. Zeocin, not Zeomycin.

Aaron

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Feb 19, 2016, 10:00:02 PM2/19/16

to Genome Engineering using CRISPR/Cas Systems

Hi Anita,

I believe the Zeocin is the "remains" of the parental vector. This is actually a SV40 em7 hybrid promoter which allows you to select for Zeocin in both mammalian cells and E. Coli.

This gene however is located outside the LTRs of the lentivirus hence will not be package into the lentivirus for integration. Hope this helps.

best,

Aaron

ajanssen3

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Feb 21, 2016, 2:18:58 PM2/21/16

to Genome Engineering using CRISPR/Cas Systems

Thanks, Aaron!

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