How to produce lenti-virus with lentiCRISPRv2 palsmids?

[Timothy Zhang]

According to Feng Zhang's protocol, to make lentivirus, lentiCRISPR (with sgRNA cloned) must be co-transfected into HEK293(F)T cells with the packaging plasmids pVSVg (AddGene 8454) and psPAX2 (AddGene 12260). But does anyone know what are the rations between these 3 plasmids when transfecting 293T cells? Thanks a lot!!

Timothy

[Neville Sanjana]

Hi Timothy, 

We often use a 4:3:1 ratio of lentiCRISPR:psPAX2:pVSVg. It is common to also see a 4:3:2 ratio in published lentivirus protocols. Either one works well.

Hope that helps,

Neville

On Thu, Jun 25, 2015 at 12:46 PM, Timothy Zhang <zinsse...@gmail.com> wrote:

According to Feng Zhang's protocol, to make lentivirus, lentiCRISPR (with sgRNA cloned) must be co-transfected into HEK293(F)T cells with the packaging plasmids pVSVg (AddGene 8454) and psPAX2 (AddGene 12260). But does anyone know what are the rations between these 3 plasmids when transfecting 293T cells? Thanks a lot!!

Timothy

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[Xiang Zhang]

Hi, Neville,

Thanks a lot!! I will try.

Timothy