By employing essential nucleotides from both the stem and loop of precursor-miRNA, greater specificity is achieved as to the mRNAs that are repressed. It is found that besides the seed sequence of the stem of the pre-miRNA, nucleotides in the loop affect the activity and specificity of the cursor- and the processing and binding to target mRNA. By using both sequences in the natural pre-miRNA or modified mimetics, one can screen for cellular miRNA expression, modulate cell properties with greater specificity and investigate cellular activity as to phenotype and response to external stimuli in the presence and absence of target protein expression.
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