Peptide nucleic acid in situ labeling (PNA-FISH) techniques provide an attractive alternative to conventional FISH and PRINS procedures for chromosomal in situ detection. PNA probes are synthetic DNA analogs with uncharged polyamide backbones. The PNA labeling reaction presents several advantages (specificity, rapidity, discriminating ability, short length of the PNA probes) that make it very attractive for cytogenetic purposes. The efficiency of centromeric and telomeric PNA probes has recently been demonstrated on human cell preparations. Their adaptation for human cells has allowed the development of new and fast molecular cytogenetic procedures, which can advantageously be used to complement FISH and PRINS techniques for the in situ assessment of aneuploidy.
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References
Lansdorp PM, Verwoerd NP, van de Rijke FM, Dragowska V, Little MT, Dirks RW, Raap AK, Tanke HJ (1996) Heterogeneity in telomere length of human chromosomes. Hum Mol Genet 5:685–691
Nielsen PE (2001) Peptide nucleic acid: A versatile tool in genetic diagnostics and molecular biology. Curr Opin Biotechnol 12:16–20
Nielsen PE, Egholm M (1999) An introduction to peptide nucleic acid. Curr Issues Mol Bio 1:89–104
Nielsen PE, Egholm M, Berg RH, Buchardt O (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254:1497–1500
Paulasova P, Andräo B, Diblik J, Macek M, Pellestor F (2004) The peptide nucleic acids as probes for chromosomal analysis: Application to human oocytes, polar bodies and preimplantation embryos. Mol Hum Reprod 10:467–472
Pellestor F, Andräo B, Taneja K, Williams B (2003) PNA on human sperm: A new approach for in situ aneuploidy estimation. Eur J Hum Genet 11:337–341
Ray A, Norden B (2000) Peptide nucleic acid (PNA): Its medical and biotechnical applications and promise for the future. FASEB J 14:1041–1060
Taneja KL, Chavez EA, Coull J, Lansdorp PM (2001) Multicolor fluorescence in situ hybridization with peptide nucleic acid probes for enumeration of specific chromosomes in human cells. Genes Chromosomes Cancer 30:57–63
Acknowledgments
The experiments performed to elaborate this protocol were partially supported by Ferring Pharmaceuticals and Organon France. P. Paulasova was supported by Czech fellowship VZ FNM 00000064203.
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Pellestor, F., Paulasova, P., Hamamah, S. (2009). PNA-FISH Technique for In Situ Assessment of Aneuploidy. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH) — Application Guide. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-70581-9_5
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