( ng protected fragment which is 4 nt lo...)Detecting miRNA & siRNA

Figure 3. Single and Multiple Target Detection. The indicated target RNAs were detected in 500, 250, 100 and 50 ng of total RNA from mouse tissues (3.5 hours exposure) or HeLa cells (6 hours exposure) as described in Figures 1 and 2. The probe specific for GAPDH mRNA (39 nt) produces a 29 nt long protected fragment with the same specific activity as the mir-16 protected fragment. miR-16 was detected with the mir-16 +4 probe.

Analysis of miRNA Expression Patterns Across Various Tissue Types
The function of most miRNAs is not known; however, a number of miRNAs seem to be involved in post-transcriptional gene regulation. Some of these miRNAs (e.g. lin-4 and let-7) inhibit protein synthesis by binding to the 3' untranslated region of target mRNAs. Others bind to perfectly complementary mRNA sequences to destroy target transcripts (e.g. Scarecrow miRNA in plants). These miRNAs, therefore, function like siRNAs and could be classified as such. Bantam, lin-4 and let-7 have also been shown to play critical roles in tissue development. Other miRNAs are believed to have similar functions because of their differential spatial and temporal expression patterns.

We used the solution hybridization assay to monitor the differential expression of two different miRNAs across mouse tissues (Figure 4). High levels of mir-16 expression were detected in all five tissues tested,
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