Human phosphoglycerate kinase 1 (PGK1) promoter

Characteristics

450 bp of 5'-flanking sequence plus the leader, first exon, and part of intron 1 of the human PGK1 gene, first sequenced by Singer-Sam et al. (1984); the fragment commonly used to drive expression spans approximately positions -528 to -13 relative to the ATG (Kita-Matsuo et al. 2009). Drives ubiquitous, moderate-level constitutive expression; in lentivirally transduced human embryonic stem cell-derived cardiomyocytes and progenitors, unsorted populations show transgene silencing that progressively drops the fraction of fluorescent-positive cells from >80% to <5% by passage 6, requiring FACS enrichment or clonal selection for stable long-term reporter lines (Kita-Matsuo et al. 2009).

Applications: Drives H2B-fluorescent reporter fusions and drug-resistance selection cassettes (Blasticidin, Neomycin, Puromycin) in lentiviral vectors for tracking and purifying human embryonic stem cell-derived cardiomyocytes, yielding >96% pure cardiomyocyte populations after dual selection (Kita-Matsuo et al. 2009); also used to drive selectable markers and Cre recombinase in lentiviral and AAV vectors.

Limitations: Unsorted lentiviral populations progressively silence PGK-driven transgene expression, falling from >80% to <5% fluorescent-positive cells by passage 6 without FACS enrichment or clonal selection (Kita-Matsuo et al. 2009); weaker than strong viral or CAG-type promoters when maximal expression level is required.

References

1. Kita-Matsuo et al. (2009). Lentiviral Vectors and Protocols for Creation of Stable hESC Lines for Fluorescent Tracking and Drug Resistance Selection of Cardiomyocytes. PLoS ONE - Kita-Matsuo 2009 hESC Cardiomyocyte