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The miRNAs are approximately ∼23 nucleotides non-coding RNAs. They regulate major processes such as development, apoptosis, cell proliferation, hematopoiesis, and patterning of the nervous system. MiRNA inhibition using antisense oligonucleotide is an effective tool for understanding miRNA function. The peptide nucleic acid (PNA) is an artificial oligonucleotide with a peptide backbone. PNAs have a stronger affinity and greater specificity than DNA or RNA and are resistant to nuclease, which is essential characteristic for a miRNA inhibitor that will be exposed to abundant serum and cellular nucleases. In here, we evaluated PNAs™ miRNA inhibitors in cultured cells including stem cell using luciferase assay and qPCR. FACS analysis was showed that PNAs™ miRNA inhibitors can penetrate to intra cells without transfection reagent. Also, miRNA inhibition effects are sustained until 15 days after treatment. PNAs™ miRNA inhibitor has a lower cytotoxicity than other nucleic acid-based miRNA inhibitors. PNAs™ miRNA inhibitors can be stored at room temperature for 20 weeks without losing the activity. These results demonstrate that PNAs are a powerful and effective tool to studies of miRNA mechanism. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4073.