Antisense Nucleic Acids in Biotechnology

Abstract

Antisense nucleic acids are natural or synthetic polymers that base pair with target RNAs and inhibit target functions. Natural antisense RNAs are found in both prokaryotic and eukaryotic organisms and regulate gene expression during cell growth or development. Synthetic and vector‐encoded antisense nucleic acids are used in experimental molecular genetics to determine the function of target genes, and are being developed for use in human disease management and in agriculture.

Keywords: antisense polynucleotides; antisense drugs; biotechnology; control of gene expression

Figure 1.

Binding of an antisense molecule to a target messenger RNA (mRNA) inhibits translation into protein. (a) Normal protein synthesis. (b) Protein synthesis inhibited by base pairing of the antisense molecule to the mRNA.

Figure 2.

Classes of antisense oligonucleotides. Base represents adenine, guanine, cytosine or thymine for oligodeoxynucleotides and uracil instead of thymine for the 2′‐O‐methyl oligoribonucleotide derivative. Peptide nucleic acids (PNAs) have adenine, guanine, cytosine and thymine or uracil as the base.

Figure 3.

Nucleic acid enzymes showing target RNA cleavage sites. (a) Schematic of hammerhead ribozyme showing binding to target RNA via Watson–Crick base pairing. Dots represent A, G, C or U. X represents A, C or U. (b) DNA enzyme binding to target RNA via Watson–Crick base pairing. Dots represent A, G, C and T for DNA enzyme and A, G, C and U for target RNA.

close

References

Agrawal S and Kandimalla ER (2000) Antisense therapeutics: is it as simple as complementary base recognition? Molecular Medicine Today 6: 72–81.

Belikova AM, Zarytova VF and Grineva NI (1967) Synthesis of ribonucleosides and diribonucleoside phosphates containing 2‐chloroethylamine and nitrogen mustard residues. Tetrahedron Letters 37: 3557–3562.

Branch AD (1998) A good antisense molecule is hard to find. Trends in Biochemical Sciences 23: 45–50.

Cai G, Zhen X, Uryu K and Friedman E (2000) Activation of extracellular signal‐regulated protein kinases is associated with a sensitized locomotor response to D(2) dopamine receptor stimulation in unilateral 6‐hydroxydopamine‐lesioned rats. Journal of Neuroscience 20: 1849–1857.

Coleman J, Green PJ and Inouye M (1984) The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes. Cell 37: 429–436.

Delihas N, Rokita SE and Zheng P (1997) Natural antisense RNA/target RNA interactions: possible models for antisense oligonucleotide drug design. Nature Biotechnology 15: 751–753.

Dias N, Dheur S, Nielsen PE et al. (1999) Antisense PNA tridecamers targeted to the coding region of Ha‐ras mRNA arrest polypeptide chain elongation. Journal of Molecular Biology 294: 403–416.

Feng et al. (1995) The RNA component of human telomerase. Science 269: 1236–1241.

Giannini CD, Roth WK, Piiper A and Zeuzem S (1999) Enzymatic and antisense effects of a specific anti‐Ki‐ras ribozyme in vitro and in cell culture. Nucleic Acids Research 27: 2737–2744.

Hoffmann B, LaPaglia SK, Kubler E et al. (2000) Developmental and metabolic regulation of the phosphoglucomutase‐encoding gene, pgmB, of Aspergillus nidulans. Molecular and General Genetics 262: 1001–1011.

Lichtsteiner SP, Lebkowski JS and Vasserot AP (1999) Telomerase. A target for anticancer therapy. Annals of the New York Academy of Sciences 886: 1–11.

Mizuno T, Chou MY and Inouye M (1984) A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proceedings of the National Academy of Sciences of the USA 81: 1966–1970.

Milner N, Mir KU and Southern EM (1997) Selecting effective antisense reagents on combinatorial oligonucleotide arrays. Nature Biotechnology 15: 537–541.

Park YG, Nesterova M, Agrawal S and Cho‐Chung YS (1999) Dual blockade of cyclic AMP response element‐ (CRE) and AP‐1‐directed transcription by CRE‐transcription factor decoy oligonucleotide. Gene‐specific inhibition of tumor growth. Journal of Biological Chemistry 274: 1573–1580.

Paterson BM, Roberts BE and Kuff EL (1977) Structural gene identification and mapping by DNA‐mRNA hybrid‐arrested cell‐free translation. Proceedings of the National Academy of Sciences of the USA 74: 4370–4374.

Pestka S, Daugherty BL, Jung V, Hotta K and Pestka RK (1984) Anti‐mRNA: specific inhibition of translation of single mRNA molecules. Proceedings of National Academy of Sciences of the USA 81: 7525–7528.

Pitts AE and Corey DR (1998) Inhibition of human telomerase by 2′‐O‐methyl‐RNA. Proceedings of the National Academy of Sciences of the USA 95: 11549–11554.

Sei S, Yang QE, O'Neill D et al. (2000) Identification of a key target sequence to block human immunodeficiency virus type 1 replication within the gag‐pol transframe domain. Journal of Virology 74: 4621–4633.

Sioud M and Leirdal M (2000) Design of nuclease resistant protein kinase c alpha DNA enzymes with potential therapeutic application. Journal of Molecular Biology 296: 937–947.

Stein CA (1999) Keeping the biotechnology of antisense in context. Nature Biotechnology 17: 209.

Stenton et al. (2000) Aerosolized sky antisense suppresses sky expression, mediator release from macrophages, and pulmonary inflammation. Journal of Immunology 164: 3790–3797.

Szklarczyk AW and Kaczmarek L (1999) Brain as a unique antisense environment. Antisense Nucleic Acid Drug Development 9: 105–116.

Tomizawa J, Itoh T, Selzer G and Som T (1981) Inhibition of ColE1 RNA primer formation by a plasmid‐specified small RNA. Proceedings of the National Academy of Sciences of the USA 78: 1421–1425.

Townsend PA, Villanova I, Uhlmann E et al. (2000) An antisense oligonucleotide targeting the alphaV integrin gene inhibits adhesion and induces apoptosis in breast cancer cells. European Journal of Cancer 36: 397–409.

Yamaoka K, Mishima K, Nagashima Y et al. (2000) Expression of galectin‐1 mRNA correlates with the malignant potential of human gliomas and expression of antisense galectin‐1 inhibits the growth of 9 glioma cells. Journal of Neuroscience Research 59: 722–730.

Wang H, Cai Q, Zeng X et al. (1999) Antitumor activity and pharmacokinetics of a mixed‐backbone antisense oligonucleotide targeted to the RI alpha subunit of protein kinase A after oral administration. Proceedings of the National Academy of Sciences of the USA 96: 13989–13994.

Zamecnik PC and Stephenson ML (1978) Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proceedings of National Academy of Sciences of the USA 75: 280–284.

Further Reading

Agrawal S and Kandimalla ER (2000) Antisense therapeutics: is it as simple as complementary base recognition? Molecular Medicine Today 6: 72–81.

Branch AD (1998) A good antisense molecule is hard to find. Trends in Biochemical Sciences 23: 45–50.

Crooke ST (2000) Progress in antisense technology: the end of the beginning. Methods in Enzymology 313: 3–45.

Finkel E (1999) DNA cuts its teeth‐as an enzyme. Science 286: 2441–2442.

Hogrefe RI (1999) An antisense oligonucleotide primer. Antisense Nucleic Acid Drug Development 9: 351–357.

Kuss B and Cotter F (1999) Antisense – time to shoot messenger. Annals of Oncology 10: 495–502.

Stein CA and Krieg AM (eds) (1998) Applied Antisense Oligonuceotide Technology. New York: Wiley‐Liss.

Szklarczyk AW and Kaczmarek L (1999) Brain as a unique antisense environment. Antisense Nucleic Acid Drug Development 9: 105–116.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Delihas, Nicholas(Apr 2001) Antisense Nucleic Acids in Biotechnology. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0000984]