pHR-CMV-TetO2-VSV-G

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The lentivirus-containing supernatant is then, normally without concentration, used to transduce the HEK293 target cell line (Fig. 1A). After genomic integration of the proviral DNA and optional enrichment using cell sorting, these cells are expanded for large-scale protein expression. The procedure can be broken down into four stages (Fig. 1B).

Protocol tips
The lentivirus-containing supernatant is then, normally without concentration, used to transduce the HEK293 target cell line (Fig. 1A). After genomic integration of the proviral DNA and optional enrichment using cell sorting, these cells are expanded for large-scale protein expression. The procedure can be broken down into four stages (Fig. 1B).

Publication protocol

The procedure to transduce expression cells is straightforward; a lentivirus producer cell line (HEK293 Lenti-X) is transiently co-transfected with transfer (encoding the transgene of interest), envelope and packaging plasmids to generate lentiviral particles. The lentivirus-containing supernatant is then, normally without concentration, used to transduce the HEK293 target cell line (Fig. 1A). After genomic integration of the proviral DNA and optional enrichment using cell sorting, these cells are expanded for large-scale protein expression. The procedure can be broken down into four stages (Fig. 1B). In the first stage (Steps 1-8 and Steps 11-14), the lentivirus producer cell line is transfected, and lentiviral particles encoding the transgene(s) are collected. In the second stage (Steps 9-10 and Steps 15-21), expression cells are infected, stably transduced, and expanded. In the third stage (Step 22 Option A i-xiii) adherent expression cells are expanded for large-scale soluble secreted protein expression. Alternatively, in the fourth stage (Step 22 Option B i-xxii), suspension expression cells are expanded for large-scale inducible membrane protein expression.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - HEK293 VSV-G using pHR-CMV-TetO2-VSV-G from Christian Siebold, Division of Structural Biology, Wellcome Cent.

Manufacturer protocol

Download the product protocol from Christian Siebold, Division of Structural Biology, Wellcome Cent for pHR-CMV-TetO2-VSV-G below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - HEK293 VSV-G using pHR-CMV-TetO2-VSV-G from Christian Siebold, Division of Structural Biology, Wellcome Cent. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.