Peptide nucleic acid (PNA) is one of the most useful DNA analogs in a wide variety of gene analysis in human cells. In order to exhibit its maximal functions, PNA must be localized to a desired place (e.g., nucleus, cytoplasm and other organelles). Here, we introduced PNAs into HeLa cells by electroporation and examined their localization at various time points. The PNA which binds to the mitochondrial COII gene was initially accumulated in the nucleus, and thereafter mostly transferred to cytoplasm. This time-dependent intracellular localization of PNA is ascribed to the breakdown of the nuclear envelope in the cell division. On the other hand, another PNA that binds to telomere repeat sequence mostly remained in the nucleus, even after the cell division occurred. The retention of this PNA in the nucleus was further enhanced when it was conjugated with Cy3.
NielsenPE, EgholmM, BergRH, BuchardtO. (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science, 254, 1497–1500.
JensenKK, OrumH, NielsenPE, NordénB. (1997) Kinetics for hybridization of peptide nucleic acids (PNA) with DNA and RNA studied with the BIAcore technique. Biochemistry, 36, 5072–5077.
4.
KnudsenH, NielsenPE. (1996) Antisense properties of duplex- and triplex-forming PNAs. Nucleic Acids Reseach, 24, 494–500.
5.
DiasN, DheurS, NielsenPE, GryaznovS, Van AerschotA, HerdewijnP, HélèneC, Saison-BehmoarasTE. (1999) Antisense PNA tridecamers targeted to the coding region of Ha-ras mRNA arrest polypeptide chain elongation. Journal of Molecular Biology, 294, 403–416.
ShiraishiT, HamzaviR, NielsenPE. (2008) Subnanomolar antisense activity of phosphonate-peptide nucleic acid (PNA) conjugates delivered by cationic lipids to HeLa cells. Nucleic Acids Research, 36, 4424–4432.
8.
IvanovaGD, ArzumanovA, AbesR, YinH, WoodMJ, LebleuB, GaitMJ. (2008) Improved cell-penetrating peptide-PNA conjugates for splicing redirection in HeLa cells and exon skipping in mdx mouse muscle. Nucleic Acids Research, 36, 6418–6428.
9.
JanowskiBA, KaihatsuK, HuffmanKE, SchwartzJC, RamR, HardyD, MendelsonCR, CoreyDR. (2005) Inhibiting transcription of chromosomal DNA with antigene peptide nucleic acids. Nature Chemical Biology, 1, 210–215.
10.
MolenaarC, WiesmeijerK, VerwoerdNP, KhazenS, EilsR, TankeHJ, DirksRW. (2003) Visualizing telomere dynamics in living mammalian cells using PNA probes. The EMBO Journal, 22, 6631–6641.
11.
ChinJY, KuanJY, LonkarPS, KrauseDS, SeidmanMM, PetersonKR, NielsenPE, KoleR, GlazerPM. (2008) Correction of a splice-site mutation in the beta-globin gene stimulated by triplex-forming peptide nucleic acids. Proceedings of the National Academy of Sciences of the United States of America, 105, 13514–13519.
12.
ItoK, KatadaH, ShigiN, KomiyamaM. (2009) Site-selective scission of human genome by artificial restriction DNA cutter. Chemical Communications, 43, 6542–6544.
13.
KatadaH, ChenHJ, ShigiN, KomiyamaM. (2009) Homologous recombination in human cells using artificial restriction DNA cutter. Chemical Communications, 43, 6545–6547.
14.
TurnerJH, RaymondJR. (2005) Interaction of calmodulin with the serotonin 5-hydroxytryptamine2A receptor. A putative regulator of G protein coupling and receptor phosphorylation by protein kinase C. The Journal of Biological Chemistry, 280, 30741–30750.
15.
TonelliR, PurgatoS, CamerinC, FronzaR, BolognaF, AlboresiS, FranzoniM, CorradiniR, SforzaS, FacciniA, ShohetJM, MarchelliR, PessionA. (2005) Anti-gene peptide nucleic acid specifically inhibits MYCN expression in human neuroblastoma cells leading to cell growth inhibition and apoptosis. Molecular Cancer Therapeutics, 4, 779–786.
16.
Ruiz-PesiniE, LottMT, ProcaccioV, PooleJC, BrandonMC, MishmarD, YiC, KreuzigerJ, BaldiP, WallaceDC. (2007) An enhanced MITOMAP with a global mtDNA mutational phylogeny. Nucleic Acids Research, 35, D823–828.
17.
MuZM, LeXF, VallianS, GlassmanAB, ChangKS. (1997) Stable overexpression of PML alters regulation of cell cycle progression in HeLa cells. Carcinogenesis, 18, 2063–2069.
