Abstract
The development of general chemical approaches for controlling gene expression would facilitate investigations into the details of cellular function (1). Antisense oligonucleotides have been widely used to block the expression of genes or alter splicing (2), but their use has suffered from questions regarding their potency and specificity. In addition, successful antisense experiments often require a large effort to identify oligomers with adequate activity (3). Peptide nucleic acids (PNAs) bind with exceptionally high affinity to complementary sequences and possess an uncharged backbone that is unlikely to prompt interactions with the many cellular proteins that bind anionic macromolecules (4). It is reasonable to hypothesize, therefore, that PNAs may possess advantages as antisense agents. In addition, PNAs have an unmatched ability to invade duplex DNA, suggesting that they may bind to chromosomal targets and act as antigene agents (4).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Stockwell B. R. (2000) Frontiers in chemical genetics. Trends Biotech. 18, 449–455.
Koller E., Gaarde W. A., and Monia B. P. (2000) Elucidating cell signalling mechanisms using antisense technology. Trends Pharm. Sci. 21, 142–148.
Stein C. A. (1999) Keeping the biotechnology of antisense in context. Nature Biotechnol. 17, 209.
Nielsen P. E., Egholm M., Berg R. H., and Buchardt O. (1991) Sequence-selective recognition of double stranded DNA by a thyminesubstituted polyamide. Science 254, 1497–1500.
Herbert B.-S., Pitts A. E., Baker S. I., Hamilton S. E., Wright W. E., Shay J. W., and Corey D. R. (1999) Inhibition of telomerase in immortal human cells leads to progressive telomere shortening and cell death. Proc. Nat. Acad. Sci. USA 96, 14,726–14,281.
Hamilton S. E., Simmons C. G., Kathriya I., and Corey D. R. (1999) Cellular delivery of peptide nucleic acids and inhibition of human telomerase. Chem. Biol. 6, 343–351.
Doyle D. F., Braasch D. A, Simmons C. G., Janowski B. A., and Corey D. R. (2001) Intracellular delivery and inhibition of gene expression by peptide nucleic acids. Biochemistry 40, 53–64.
Shammas M. A., Simmons C. G., Corey D. R., and Shmookler-Reis R. J. (1999) Inhibition of telomerase reverses immortality of transformed. Oncogene 18, 6191–6200.
Faruqi A. F., Egholm M., and Glazer P. M. (1997) Peptide nucleic acid targeted mutagenesis of a chromosomal gene in mouse cells. Proc. Natl. Acad. Sci. USA 95, 1398–1403.
Simmons C. G., Pitts A. E., Mayfield L. D., Shay J. W., and Corey D. R. (1997) Synthesis and membrane permeability of PNA-peptide conjugates. Bioorg. Med. Chem. Lett. 7, 3001–3007.
Pooga M., Soomets U., Hallbrink M., Valkna A., Saar K., Kahl U., et al. (1998) Cell penetrating PNA constructs regulate galanin receptor levels and modify pain transmission in vivo. Nature Biotech. 16, 857–861.
Mayfield L. D. and Corey D. R. (1999) Automated synthesis of peptide nucleic acids (PNAs) and peptide nucleic acid-peptide conjugates Anal. Biochem. 268, 401–404.
Norton J. C., Waggenspack J. J., Varnum E., and Corey D. R. (1995) Targeting peptide nucleic acid protein conjugates to structural features within duplex DNA. Bioorg. Med. Chem. 3, 437–445.
Goodwin T. E., Holland R. D., Lay J. O., and Raney K. D. (1998) A simple procedure for solid-phase synthesis of peptide nucleic acids with N-terminal cysteine. Bioorg. Med. Chem. Lett. 8, 2231–2234.
Mayfield L. D. and Corey D. R. (1999) Enhancing solid phase synthesis by a noncovalent protection strategy: efficient coupling of rhodamine to peptide nucleic acids. Bioorganic Med. Chem. Lett. 9, 1419–1422.
Atkins P. W. (1990) Physical Chemistry, 4th ed. W. H. Freeman and Company, New York, p. 219.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Braasch, D.A., Corey, D.R. (2002). Lipid-Mediated Introduction of Peptide Nucleic Acids Into Cells. In: Nielsen, P.E. (eds) Peptide Nucleic Acids. Methods in Molecular Biology, vol 208. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-290-2:211
Download citation
DOI: https://doi.org/10.1385/1-59259-290-2:211
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-976-6
Online ISBN: 978-1-59259-290-6
eBook Packages: Springer Protocols