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. 2016 Oct 1;27(10):847–859. doi: 10.1089/hum.2016.065

Figure 1.

Figure 1.

MicroRNA-126 (miRNA126) target elements in lentiviral (LV) vectors restrict transgene expression in human CD34+ hematopoietic stem and progenitor cells but allow robust transgene expression in natural killer (NK) cells. (a) Bidirectional lentiviral vector design with a mini-cytomegalovirus (CMV) promoter driving a truncated nerve growth factor receptor (tNGFR) cDNA in one direction and phosphoglycerate kinase (PGK) promoter driving enhanced green fluorescent protein (eGFP) cDNA with (4T) or without (0T) four repeats of the miR126 target sequences in the opposite direction. (b) Human CD34+ cells were transduced with the two LV vectors. Analysis of tNGFR and eGFP expression by flow cytometry 36 hr posttransduction (left); and after 18 days of NK (CD56+) cell differentiation (right) is shown. The cells expressing tNGFR or eGFP are gated on the basis of appropriate negative controls, and their percentage is indicated in each dot plot. (c) Cumulative data on the proportion of transduced CD34+ cells (tNGFR+; blue columns) that are expressing GFP (green columns) 36 hr after transduction or after 18 days of NK cell differentiation with the 0T and 4T LV vectors. Error bars represent the SEM. Statistics were done by two-way analysis of variance with Sidak correction. ****p < 0.0001.