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The let-7 microRNA Family Regulates RAS: Implications for Development and Oncogenesis

n, RAS expression was monitored by immunofluorescence (A, C) and quantified using MetaMorph software (Universal Imaging corporation) (B, D; triplicate assays). Modified with permission from Cell 120: 635647.


In reciprocal experiments, HeLa cells, which express high levels of let-7a, were transfected with a synthetic, antisense RNA to let-7a (let-7a Anti-miR miRNA Inhibitor), and the prediction that inhibiting let-7 activity may de-repress RAS expression held true (Figures 1CD). RAS protein levels were ~70% higher in cells transfected with the let-7a inhibitor compared to cells transfected with a negative control. Together, these results suggest that let-7 negatively regulates the expression of RAS in human cells.


let-7 regulates the Human RAS genes via their 3'UTRs

Portions of the NRAS 3'UTR (3.5 kb containing nine LCSs) and the KRAS 3'UTR (1 kb containing seven LCSs) were individually subcloned downstream of a luciferase reporter. When transfected into HeLa cells, the reporter constructs with the RAS 3'UTRs expressed 4- to 8-fold less luciferase activity than the control reporter construct with no insert (Figure 2A).

Figure 2. let-7 Regulates NRAS and KRAS Through Their 3' UTRs. The NRAS 3'UTR (3.5 kb containing nine LCSs) and the KRAS 3'UTR (1 kb containing seven LCSs) were subcloned downstream of t
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