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miRNA Expression in White Blood Cells

ed from the fractionated white blood cells was quantitated by UV absorbance and analyzed by ethidium bromide staining on a 15% denaturing polyacrylamide gel. RNA yields ranged from about 20-40 g per 10 ml blood sample. No obvious degradation of the ribosomal RNA and tRNA bands was observed. Four representative samples are presented in Figure 2.

Figure 2. Analysis of miRNA Expression in White Blood Cells. Leukocytes were collected from 10 ml EDTA-anticoagulated blood samples from four different donors, and total RNA was isolated with the mirVana miRNA Isolation Kit. Samples were analyzed as described in Figure 1 except that a probe specific for miR-22 miRNA was used in the solution hybridization assay.


The RNA samples were then analyzed for the presence of several small RNAs using Northern blot or the mirVana miRNA Detection Kit. Interestingly, several miRNAs--let-7, miR-16, and miR-22--were detected in total RNA from white blood cells (Figure 2). miR-16 was the most abundant miRNA in all of the 14 samples. A direct comparison of total RNA samples from other human tissues showed that miR-16 is expressed at a higher level in white blood cells than in any other tissues we have examined (data not shown). While different expression levels were observed across donors for the few miRNA tested, there was no clear donor-specific difference in the global miRNA expression pattern. Experiments are in progress to determine whether other miRNAs may have donor-specific expression patterns and whether these patterns
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