BioPharmaNet, Department of Biochemistry and Molecular Biology, University of Ferrara, via Fossato di Mortara 74, 44100 Ferrara, Italy
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, University of Ferrara, Ferrara, Italy
Institute of Biostructure and Bioimaging, University of Naples, Naples, Italy
School of Biotechnological Sciences, University of Naples Federico II, Naples, Italy
References: Nielsen, P.E., Egholm, M., Berg, R.H., Buchardt, O., Sequence selective recognition of DNA by strand displacement with a thymine-substituted polyamide (1991) Science, 254, pp. 1497-150
Lundin, K.E., Good, L., Strömberg, R., Gräslund, A., Smith, C.I., Biological activity and biotechnological aspects of peptide nucleic acid (2006) Adv Genet, 56, pp. 1-51
Nielsen, E., RNA targeting using peptide nucleic acid (2006) Handb Exp Pharmacol, 173, pp. 395-403
Yin, H., Lu, Q., Wood, M., Effective exon skipping and restoration of dystrophin expression by peptide nucleic acid antisense oligonucleotides in mdx mice (2008) Mol Ther, 16, pp. 38-45
Romanelli, A., Pensato, S., Di Niola, E., Feriotto, G., Breveglieri, G., Zaccaro, L., Antisense PNA and PNA-peptide conjugates for the modulation of the β-globin gene splicing (2005) Biopolymers, 80, pp. 551-552
Nielsen, P.E., Gene targeting using peptide nucleic acid (2005) Methods Mol Biol, 288, pp. 343-358
Tonelli, R., Purgato, S., Camerin, C., Fronza, R., Bologna, F., Alboresi, S., Anti-gene peptide nucleic acid specifically inhibits MYCN expression in human neuroblastoma cells leading to cell growth inhibition and apoptosis (2005) Mol Cancer Ther, 4, pp. 779-786
Pesce, C.D., Bolacchi, F., Bongiovanni, B., Cisotta, F., Capozzi, M., Diviacco, S., Anti-gene peptide nucleic acid targeted to proviral HIV-1 DNA inhibits in vitro HIV-1 replication (2005) Antiviral Res, 66, pp. 13-22
Corradini, R., Feriotto, G., Sforza, S., Marchelli, R., Gambari, R., Enhanced recognition of cystic fibrosis W1282X DNA point mutation by chiral peptide nucleic acid probes by a surface plasmon resonance biosensor (2004) J Mol Recognit, 17, pp. 76-84
Feriotto, G., Corradini, R., Sforza, S., Bianchi, N., Mischiati, C., Marchelli, R., Peptide nucleic acids and biosensor technology for realtime detection of the cystic fibrosis W1282X mutation by surface plasmon resonance (2001) Lab Invest, 81, pp. 1415-1427
Ohtsuki, T., Fujimoto, T., Kamimukai, M., Kumano, C., Kitamatsu, M., Sisido, M., Isolation of small RNAs using biotinylated PNAs (2008) J Biochem, 144, pp. 415-418
Bonham, M.A., Brown, S., Boyd, A.L., Brown, P.H., Bruckenstein, D.A., Hanvey, J.C., An assessment of the antisense properties of RNase H-competent and steric-blocking oligomers (1995) Nucleic Acids Res, 23, pp. 1197-1203
Weatherall, D.J., Clegg, J.B., (2001) The thalassemia syndromes, pp. 133-191. , Oxford: Blackwell Scientific
Ho, P.J., The regulation of beta globin gene expression and beta thalassemia (1999) Pathology, 31, pp. 315-324
Eleftheriou, A., The pathophysiology of thalassaemia: Lack of β-chains and excess of α-chains (2007) About Thalassemia, pp. 16-18. , Eleftheriou A, editor, Nicosia: The Thalassemia International Federation publisher
Schrier, S.L., Pathophysiology of thalassemia (2002) Curr Opin Hematol, 9, pp. 123-126
Bank, A., Regulation of human fetal hemoglobin: New players, new complexities (2006) Blood, 107, pp. 435-443
Voon, H.P., Wardan, H., Vadolas, J., Co-inheritance of alpha- and beta-thalassaemia in mice ameliorates thalassaemic phenotype (2007) Blood Cells Mol Dis, 39, pp. 184-188
Gambari, R., Fibach, E., Medicinal chemistry of fetal hemoglobin inducers for treatment of beta-thalassemia (2007) Curr Med Chem, 14, pp. 199-212
Xie, S.Y., Ren, Z.R., Zhang, J.Z., Guo, X.B., Wang, Q.X., Wang, S., Restoration of the balanced alpha/beta-globin gene expression in beta654-thalassemia mice using combined RNAi and antisense RNA approach (2007) Hum Mol Genet, 16, pp. 2616-2625
Bianchi, N., Borgatti, M., Gambari, R., Gene Silencing in thalassemia (2008) Minerva Biotecnologica, 20, pp. 51-55
Abes, S., Ivanova, G.D., Abes, R., Arzumanov, A.A., Williams, D., Owen, D., Peptide-based delivery of steric-block PNA oligonucleotides (2009) Methods Mol Biol, 480, pp. 85-99
Lundin, P., Johansson, H., Guterstam, P., Holm, T., Hansen, M., Langel, U., Distinct uptake routes of cell-penetrating peptide conjugates (2008) Bioconjug Chem, 19, pp. 2535-2542
Ivanova, G.D., Arzumanov, A., Abes, R., Yin, H., Wood, M.J., Lebleu, B., Improved cell-penetrating peptide-PNA conjugates for splicing redirection in HeLa cells and exon skipping in mdx mouse muscle (2008) Nucleic Acids Res, 36, pp. 6418-6428
Ivanova, G.D., Fabani, M.M., Arzumanov, A.A., Abes, R., Yin, H., Lebleu, B., PNA-peptide conjugates as intracellular gene control agents (2008) Nucleic Acids Symp Ser (Oxf), 52, pp. 31-32
Wright, D.G., Zhang, Y., Murphy, J.R., Effective delivery of antisense peptide nucleic acid oligomers into cells by anthrax protective antigen (2008) Biochem Biophys Res Commun, 376, pp. 200-205
Shiraishi, T., Hamzavi, R., Nielsen, P.E., Subnanomolar antisense activity of phosphonate-peptide nucleic acid (PNA) conjugates delivered by cationic lipids to HeLa cells (2008) Nucleic Acids Res, 36, pp. 4424-4432
Robertson, K.L., Yu, L., Armitage, B.A., Lopez, A.J., Peteanu, L.A., Fluorescent PNA probes as hybridization labels for biological RNA (2006) Biochemistry, 45, pp. 6066-6074
Rasmussen, F.W., Bendifallah, N., Zachar, V., Shiraishi, T., Fink, T., Ebbesen, P., Evaluation of transfection protocols for unmodified and modified peptide nucleic acid (PNA) oligomers (2006) Oligonucleotides, 16, pp. 43-57
Albertshofer, K., Siwkowski, A.M., Wancewicz, E.V., Esau, C.C., Watanabe, T., Nishihara, K.C., Structure-activity relationship study on a simple cationic peptide motif for cellular delivery of antisense peptide nucleic acid (2005) J Med Chem, 48, pp. 6741-6749
Karras, J.G., Maier, M.A., Lu, T., Watt, A., Manoharan, M., Peptide nucleic acids are potent modulators of endogenous pre-mRNA splicing of the murine interleukin-5 receptor-alpha chain (2001) Biochemistry, 40, pp. 7853-7859
Siwkowski, A.M., Malik, L., Esau, C.C., Maier, M.A., Wancewicz, E.V., Albertshofer, K., Identification and functional validation of PNAs that inhibit murine CD40 expression by redirection of splicing (2004) Nucleic Acids Res, 32, pp. 2695-2706
Wilusz, J.E., Devanney, S.C., Caputi, M., Chimeric peptide nucleic acid compounds modulate splicing of the bcl-x gene in vitro and in vivo (2005) Nucleic Acids Res, 33, pp. 6547-6554
El-Andaloussi, S., Johansson, H.J., Lundberg, P., Langel, U., Induction of splice correction by cell-penetrating peptide nucleic acids (2006) J Gene Med, 8, pp. 1262-1273
van Vliet, L., de Winter, C.L., van Deutekom, J.C., van Ommen, G.J., Aartsma-Rus, A., Assessment of the feasibility of exon 45-55 multiexon skipping for Duchenne muscular dystrophy (2008) BMC Med Genet, 9, pp. 105-111
Gao, Y., Zu, T., Low, W.C., Orr, H.T., McIvor, R.S., Antisense RNA sequences modulating the ataxin-1 message: Molecular model of gene therapy for spinocerebellar ataxia type 1, a dominant-acting unstable trinucleotide repeat disease (2008) Cell Transplant, 17, pp. 723-734
Davis, R.L., Homer, V.M., George, P.M., Brennan, S.O., A deep intronic mutation in FGB creates a consensus exonic splicing enhancer motif that results in afibrinogenemia caused by aberrant mRNA splicing, which can be corrected in vitro with antisense oligonucleotide treatment (2009) Hum Mutat, 30, pp. 221-227
Madsen, E.C., Morcos, P.A., Mendelsohn, B.A., Gitlin, J.D., In vivo correction of a Menkes disease model using antisense oligonucleotides (2008) Proc Natl Acad Sci USA, 105, pp. 3909-3914
Mascellani, N., Tagliavini, L., Gamberoni, G., Rossi, S., Marchesini, J., Taccioli, C., Using miRNA expression data for the study of human cancer (2008) Minerva Biotecnologica, 20, pp. 23-30
Habbe, N., Koorstra, J.B., Mendell, J.T., Offerhaus, G.J., Ryu, J.K., Feldmann, G., MicroRNA miR-155 is a biomarker of early pancreatic neoplasia (2009) Cancer Biol Ther, 8, pp. 25-31
Duisters, R.F., Tijsen, A.J., Schroen, B., Leenders, J.J., Lentink, V., van der Made, I., miR-133 and miR-30 regulate connective tissue growth factor: Implications for a role of microRNAs in myocardial matrix remodeling (2009) Circ Res, 104, pp. 170-178
Chen, J.F., Callis, T.E., Wang, D.Z., microRNAs and muscle disorders (2009) J Cell Sci, 122, pp. 13-20
Xiao, B., Guo, J., Miao, Y., Jiang, Z., Huan, R., Zhang, Y., Detection of miR-106a in gastric carcinoma and its clinical significance (2009) Clin Chim Acta, 400, pp. 97-102
Abes, S., Turner, J.J., Ivanova, G.D., Owen, D., Williams, D., Arzumanov, A., Efficient splicing correction by PNA conjugation to an R6-Penetratin delivery peptide (2007) Nucleic Acids Res, 35, pp. 4495-4502
Nielsen, P. E., Egholm, M., Berg, R. H., Buchardt, O., Sequence selective recognition of DNA by strand displacement with a thymine-substituted polyamide (1991) Science, 254, pp. 1497-150
Lundin, K. E., Good, L., Str mberg, R., Gr slund, A., Smith, C. I., Biological activity and biotechnological aspects of peptide nucleic acid (2006) Adv Genet, 56, pp. 1-51
Nielsen, P. E., Gene targeting using peptide nucleic acid (2005) Methods Mol Biol, 288, pp. 343-358
Pesce, C. D., Bolacchi, F., Bongiovanni, B., Cisotta, F., Capozzi, M., Diviacco, S., Anti-gene peptide nucleic acid targeted to proviral HIV-1 DNA inhibits in vitro HIV-1 replication (2005) Antiviral Res, 66, pp. 13-22
Bonham, M. A., Brown, S., Boyd, A. L., Brown, P. H., Bruckenstein, D. A., Hanvey, J. C., An assessment of the antisense properties of RNase H-competent and steric-blocking oligomers (1995) Nucleic Acids Res, 23, pp. 1197-1203
Weatherall, D. J., Clegg, J. B., (2001) The thalassemia syndromes, pp. 133-191. , Oxford: Blackwell Scientific
Ho, P. J., The regulation of beta globin gene expression and beta thalassemia (1999) Pathology, 31, pp. 315-324
Schrier, S. L., Pathophysiology of thalassemia (2002) Curr Opin Hematol, 9, pp. 123-126
Voon, H. P., Wardan, H., Vadolas, J., Co-inheritance of alpha- and beta-thalassaemia in mice ameliorates thalassaemic phenotype (2007) Blood Cells Mol Dis, 39, pp. 184-188
Xie, S. Y., Ren, Z. R., Zhang, J. Z., Guo, X. B., Wang, Q. X., Wang, S., Restoration of the balanced alpha/beta-globin gene expression in beta654-thalassemia mice using combined RNAi and antisense RNA approach (2007) Hum Mol Genet, 16, pp. 2616-2625
Ivanova, G. D., Arzumanov, A., Abes, R., Yin, H., Wood, M. J., Lebleu, B., Improved cell-penetrating peptide-PNA conjugates for splicing redirection in HeLa cells and exon skipping in mdx mouse muscle (2008) Nucleic Acids Res, 36, pp. 6418-6428
Ivanova, G. D., Fabani, M. M., Arzumanov, A. A., Abes, R., Yin, H., Lebleu, B., PNA-peptide conjugates as intracellular gene control agents (2008) Nucleic Acids Symp Ser (Oxf), 52, pp. 31-32
Wright, D. G., Zhang, Y., Murphy, J. R., Effective delivery of antisense peptide nucleic acid oligomers into cells by anthrax protective antigen (2008) Biochem Biophys Res Commun, 376, pp. 200-205
Robertson, K. L., Yu, L., Armitage, B. A., Lopez, A. J., Peteanu, L. A., Fluorescent PNA probes as hybridization labels for biological RNA (2006) Biochemistry, 45, pp. 6066-6074
Rasmussen, F. W., Bendifallah, N., Zachar, V., Shiraishi, T., Fink, T., Ebbesen, P., Evaluation of transfection protocols for unmodified and modified peptide nucleic acid (PNA) oligomers (2006) Oligonucleotides, 16, pp. 43-57
Karras, J. G., Maier, M. A., Lu, T., Watt, A., Manoharan, M., Peptide nucleic acids are potent modulators of endogenous pre-mRNA splicing of the murine interleukin-5 receptor-alpha chain (2001) Biochemistry, 40, pp. 7853-7859
Siwkowski, A. M., Malik, L., Esau, C. C., Maier, M. A., Wancewicz, E. V., Albertshofer, K., Identification and functional validation of PNAs that inhibit murine CD40 expression by redirection of splicing (2004) Nucleic Acids Res, 32, pp. 2695-2706
Wilusz, J. E., Devanney, S. C., Caputi, M., Chimeric peptide nucleic acid compounds modulate splicing of the bcl-x gene in vitro and in vivo (2005) Nucleic Acids Res, 33, pp. 6547-6554
Davis, R. L., Homer, V. M., George, P. M., Brennan, S. O., A deep intronic mutation in FGB creates a consensus exonic splicing enhancer motif that results in afibrinogenemia caused by aberrant mRNA splicing, which can be corrected in vitro with antisense oligonucleotide treatment (2009) Hum Mutat, 30, pp. 221-227
Madsen, E. C., Morcos, P. A., Mendelsohn, B. A., Gitlin, J. D., In vivo correction of a Menkes disease model using antisense oligonucleotides (2008) Proc Natl Acad Sci USA, 105, pp. 3909-3914
Duisters, R. F., Tijsen, A. J., Schroen, B., Leenders, J. J., Lentink, V., van der Made, I., miR-133 and miR-30 regulate connective tissue growth factor: Implications for a role of microRNAs in myocardial matrix remodeling (2009) Circ Res, 104, pp. 170-178
Chen, J. F., Callis, T. E., Wang, D. Z., microRNAs and muscle disorders (2009) J Cell Sci, 122, pp. 13-20
PNA-mediated alteration of mRNA splicing
PNAs are expected to be very active in altering the splicing pattern of target RNA sequences. Several reports have been published on this specific task, focused on analysis of PNA altering splicing and vehiculated with a variety of delivery systems. The conclusions, here reviewed, are that PNAs altering splicing have important biological effects. In our own laboratory, we have studied the in vitro effects of a 14-mer PNA targeting a portion of the β-globin gene intron 1, suggesting that this strategy might be of interest to counteract the excess production of β-globin. Furthermore, the data obtained on the same 14-mer PNA, conjugated to the SV40 NLS peptide (β-globin-PNA/ NLS) to enhance the PNA concentration in cells, demonstrate that the NLS peptide does not interfere with inhibition of the splicing.