pHR-CMV-TetO2-CD45

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	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.

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