Skip to main content
NCBI home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 2006;173:405–422. doi: 10.1007/3-540-27262-3_21

Locked Nucleic Acid: High-Affinity Targeting of Complementary RNA for RNomics

S Kauppinen 10, B Vester 11, J Wengel 12
Editors: Volker Erdmann7, Jan Barciszewski8, Jürgen Brosius9
PMCID: PMC7120141  PMID: 16594628

Abstract

Locked nucleic acid (LNA) is a nucleic acid analog containing one or more LNA nucleotide monomers with a bicyclic furanose unit locked in an RNA-mimicking sugar conformation. This conformational restriction is translated into unprecedented hybridization affinity towards complementary single-stranded RNA molecules. That makes fully modified LNAs, LNA/DNA mixmers, or LNA/RNA mixmers uniquely suited for mimicking RNA structures and for RNA targeting in vitro or in vivo. The focus of this chapter is on LNA antisense, LNA-modified DNAzymes (LNAzymes), LNA-modified small interfering (si)RNA (siLNA), LNA-enhanced expression profiling by real-time RT-PCR and detection and analysis of microRNAs by LNA-modified probes.

Keywords: LNA, Locked nucleic acid, RNA targeting, LNA antisense, MicroRNA targeting

Contributor Information

Volker Erdmann, Email: erdmann@chemie.fu-berlin.de.

Jan Barciszewski, Email: jan.barciszewski@ibch.poznan.pl.

Jürgen Brosius, Email: brosius@pop.uni-muenster.de.

J. Wengel, Email: jwe@chem.sdu.dk

References

  1. Ambros V. MicroRNAs: tiny regulators with great potential. Cell. 2001;107:823–826. doi: 10.1016/S0092-8674(01)00616-X. [DOI] [PubMed] [Google Scholar]
  2. Ambros V., Lee R.C., Lavanway A., Williams P.T., Jewell D. MicroRNAs and other tiny endogenous RNAs in C. elegans. Curr Biol. 2003;13:807–818. doi: 10.1016/S0960-9822(03)00287-2. [DOI] [PubMed] [Google Scholar]
  3. Arzumanov A., Stetsenko D.A., Malakhov A.D., Reichelt S., Sørensen M.D., Babu B.R., Wengel J., Gait M.J. A structure-activity study of the inhibition of HIV-1 tat-dependent transactivation by mixmer 2′-O-methyl oligoribonucleotides containing locked nucleic acid (LNA), α-L-LNA, or 2′-thio-LNA residues. Oligonucleotides. 2003;13:435–45. doi: 10.1089/154545703322860762. [DOI] [PubMed] [Google Scholar]
  4. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA. 1972;69:1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bartel D.P. MicroRNAs: genomics, biogenesis, mechanism and function. Cell. 2004;116:281–297. doi: 10.1016/S0092-8674(04)00045-5. [DOI] [PubMed] [Google Scholar]
  6. Benenson Y., Gil B., Ben-Dor U., Adar R., Shapiro E. An autonomous molecular computer for logical control of gene expression. Nature. 2004;429:423–429. doi: 10.1038/nature02551. [DOI] [PubMed] [Google Scholar]
  7. Braasch D.A., Corey D.R. Locked nucleic acid (LNA): fine-tuning the recognition of DNA and RNA. Chem Biol. 2001;8:1–7. doi: 10.1016/S1074-5521(00)00058-2. [DOI] [PubMed] [Google Scholar]
  8. Braasch D.A., Jensen S., Liu Y., Kaur K., Arar K., White M.A., Corey D.R. RNA interference in mammalian cells by chemically-modified RNA. Biochemistry. 2003;8:7967–7975. doi: 10.1021/bi0343774. [DOI] [PubMed] [Google Scholar]
  9. Bustin S.A. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000;25:169–193. doi: 10.1677/jme.0.0250169. [DOI] [PubMed] [Google Scholar]
  10. Childs J.L., Disney M.D., Turner D.H. Oligonucleotide directed misfolding of RNA inhibits Candida albicans group I intron splicing. Proc Natl Acad Sci USA. 2002;99:11091–11096. doi: 10.1073/pnas.172391199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chirgwin J.M., Przybyla A.E., MacDonald R.J., Rutter W.J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979;18:5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  12. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159. doi: 10.1016/0003-2697(87)90021-2. [DOI] [PubMed] [Google Scholar]
  13. Crinelli R., Bianchi M., Gentilini L., Magnani M. Design and characterization of decoy oligonucleotides containing locked nucleic acids. Nucleic Acids Res. 2002;30:2435–2443. doi: 10.1093/nar/30.11.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. De Giusto D.A., King G.C. Strong positional preference in the interaction of LNA oligonucleotides with DNA polymerase and proofreading exonuclease activities: implications for genotyping assays. Nucleic Acids Res. 2004;32:e32. doi: 10.1093/nar/gnh036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Elayadi A.N., Braasch D.A., Corey D.R. Implications of high-affinity hybridization by locked nucleic acid oligomers for inhibition of human telomerase. Biochemistry. 2002;41:9973–9981. doi: 10.1021/bi025907j. [DOI] [PubMed] [Google Scholar]
  16. Elmén J., Zhang H.Y., Zuber B., Ljungberg K., Wahren B., Wahlestedt C., Liang Z. Locked nucleic acid containing antisense oligonucleotides enhance inhibition of HIV-1 genome dimerization and inhibit virus replication. FEBS Lett. 2004;578:285–290. doi: 10.1016/j.febslet.2004.11.015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Elmén J., Thonberg H., Ljungberg K., Frieden M., Westergaard M., Xu Y., Wahren B., Liang Z., Ørum H., Koch T., Wahlestedt C. Locked nucleic acid (LNA) mediated improvements in siRNA stability and functionality. Nucleic Acids Res. 2005;33:439–447. doi: 10.1093/nar/gki193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fahmy R.G., Khachigian L.M. Locked nucleic acid modified DNA enzymes targeting early growth response-1 inhibit human vascular smooth muscle cell growth. Nucleic Acids Res. 2004;32:2281–2285. doi: 10.1093/nar/gkh543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fire A., Xu S., Montgomery M.K., Kostas S.A., Driver S.E., Mello C.C. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998;391:806–811. doi: 10.1038/35888. [DOI] [PubMed] [Google Scholar]
  20. Fluiter K., ten Asbroek L.M.A., de Wissel M.B., Marit B., Jakobs M.E., Wissenbach M., Olsson H., Olsen O., Oerum H., Baas F. In vivo tumor growth inhibition and biodistribution studies of locked nucleic acid (LNA) antisense oligonucleotides. Nucleic Acids Res. 2003;31:953–962. doi: 10.1093/nar/gkg185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Frieden M., Christensen S.M., Mikkelsen N.D., Rosenbohm C., Thrue C.A., Westergaard M., Hansen H.F., Ørum H., Koch T. Expanding the design horizon of antisense oligonucleotides with alpha-L-LNA. Nucleic Acids Res. 2003;31:6365–6372. doi: 10.1093/nar/gkg820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Frieden M., Hansen H.F., Koch T. Nuclease stability of LNA oligonucleotides and LNA-DNA chimeras. Nucleosides Nucleotides Nucleic Acids. 2003;22:1041–1043. doi: 10.1081/NCN-120022731. [DOI] [PubMed] [Google Scholar]
  23. Griffiths-Jones S. The microRNA registry. Nucleic Acids Res. 2004;32:D109–D111. doi: 10.1093/nar/gkh023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Grünweller A., Wyszko E., Bieber B., Jahnel R., Erdmann V.A., Kurreck J. Comparison of different antisense strategies in mammalian cells using locked nucleic acids, 2′-Omethyl RNA, phosphorothioates and small interfering RNA. Nucleic Acids Res. 2003;31:3185–3193. doi: 10.1093/nar/gkg409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hansen J.B., Westergaard M., Thrue C.A., Giwercman B., Oerum H. Antisense knockdown of PKC-alpha using LNA-oligos. Nucleosides Nucleotides Nucleic Acids. 2003;22:1607–1609. doi: 10.1081/ncn-120023045. [DOI] [PubMed] [Google Scholar]
  26. Ittig D., Liu S., Renneberg D., Schuemperli D., Leumann C.J. Nuclear antisense effects in cyclophilin A pre-mRNA splicing by oligonucleotides: a comparison of tricyclo-DNA with LNA. Nucleic Acids Res. 2004;32:346–353. doi: 10.1093/nar/gkh187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jacobsen N., Nielsen P.S., Jeffares D.C., Eriksen J., Ohlsson H., Arctander P., Kauppinen S. Direct isolation of poly(A)+ RNA from 4 M guanidine thiocyanate-lysed cell extracts using locked nucleic acid-oligo(T) capture. Nucleic Acids Res. 2004;32:e64/1–e64/10. doi: 10.1093/nar/gnh056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Jepsen J.S., Wengel J. LNA-antisense rivals siRNA for gene silencing. Curr Opin Drug Discov Devel. 2004;7:188–194. [PubMed] [Google Scholar]
  29. Kampa D., Cheng I.J., Kapranov P., Yamanaka M., Brubaker S., Cawley S., Drenkow J., Piccolboni A., Bekiranov S., Helt G., Tammana H., Gingeras T.R. Novel RNAs identified from an in-depth analysis of the transcriptome of human chromosomes 21 and 22. Genome Res. 2004;14:331–342. doi: 10.1101/gr.2094104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ke X.-S., Liu C.-M., Liu D.-P., Liang C.-C. MicroRNAs: key participants in gene regulatory networks. Curr Opin Chem Biol. 2003;7:516–523. doi: 10.1016/S1367-5931(03)00075-9. [DOI] [PubMed] [Google Scholar]
  31. Khachigian L.M., Fahmy R.G., Zhang G., Bobryshev Y.V., Kaniaros A. c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury. Inhibition by a novel DNA enzyme targeting c-Jun. J Biol Chem. 2002;277:22985–22991. doi: 10.1074/jbc.M200977200. [DOI] [PubMed] [Google Scholar]
  32. Koshkin A.A., Nielsen P., Meldgaard M., Rajwanshi V.K., Singh S.K., Wengel J. LNA (locked nucleic acid): an RNA mimic forming exceedingly stable LNA:LNA duplexes. J Am Chem Soc. 1998;120:13252–13253. doi: 10.1021/ja9822862. [DOI] [Google Scholar]
  33. Koshkin A.A., Singh S.K., Nielsen P., Rajwanshi V.K., Kumar R., Meldgaard M., Olsen C.E., Wengel J. LNA (locked nucleic acids): synthesis of the adenine, cytosine, guanine, 5-methylcytosine, thymine and uracil bicyclonucleoside monomers, oligomerisation, and unprecedented nucleic acid recognition. Tetrahedron. 1998;54:3607–3630. doi: 10.1016/S0040-4020(98)00094-5. [DOI] [Google Scholar]
  34. Kumar R., Singh S.K., Koshkin A.A., Rajwanshi V.K., Meldgaard M., Wengel J. The first analogues of LNA (locked nucleic acids): Phosphorothioate-LNA and 2′-thio-LNA. Bioorg Med Chem Lett. 1998;8:2219–2222. doi: 10.1016/S0960-894X(98)00366-7. [DOI] [PubMed] [Google Scholar]
  35. Kurreck J., Wyszko E., Gillen C., Erdmann V.A. Design of antisense oligonucleotides stabilized by locked nucleic acids. Nucleic Acids Res. 2002;30:1911–1918. doi: 10.1093/nar/30.9.1911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lagos-Quintana M., Rauhut R., Lendeckel W., Tuschl T. Identification of novel genes coding for small expressed RNAs. Science. 2001;294:853–858. doi: 10.1126/science.1064921. [DOI] [PubMed] [Google Scholar]
  37. Lander E.S., Linton L.M., Birren B., Nusbaum C., Zody M.C., Baldwin J., Devon K., Dewar K., Doyle M., FitzHugh W., Funke R., Gage D., Harris K., Heaford A., Howland J., Kann L., Lehoczky J., LeVine R., McEwan P., McKernan K., Meldrim J., Mesirov J. P., Miranda C., Morris W., Naylor J., Raymond C., Rosetti M., Santos R., Sheridan A., Sougnez C., Stange-Thomann N., Stojanovic N., Subramanian A., Wyman D., Rogers J., Sulston J., Ainscough R., Beck S., Bentley D., Burton J., Clee C., Carter N., Coulson A., Deadman R., Deloukas P., Dunham A., Dunham I., Durbin R., French L., Grafham D., Gregory S., Hubbard T., Humphray S., Hunt A., Jones M., Lloyd C., McMurray A., Matthews L., Mercer S., Milne S., Mullikin J.C., Mungall A., Plumb R., Ross M., Shownkeen R., Sims S., Waterston R.H., Wilson R.K., Hillier L.W., McPherson J.D., Marra M.A., Mardis E.R., Fulton L.A., Chinwalla A.T., Pepin K.H., Gish W.R., Chissoe S.L., Wendl M.C., Delehaunty K.D., Miner T.L., Delehaunty A., Kramer J.B., Cook L.L., Fulton R.S., Johnson D.L., Minx P.J., Clifton S.W., Hawkins T., Branscomb E., Predki P., Richardson P., Wenning S., Slezak T., Doggett N., Cheng J.F., Olsen A., Lucas S., Elkin C., Uberbacher E., Frazier M., Gibbs R.A., Muzny D.M., Scherer S.E., Bouck J.B., Sodergren E.J., Worley K.C., Rives C.M., Gorrell J.H., Metzker M.L., Naylor S.L., Kucherlapati R.S., Nelson D.L., Weinstock G.M., Sakaki Y., Fujiyama A., Hattori M., Yada T., Toyoda A., Itoh T., Kawagoe C., Watanabe H., Totoki Y., Taylor T., Weissenbach J., Heilig R., Saurin W., Artiguenave F., Brottier P., Bruls T., Pelletier E., Robert C., Wincker P., Smith D.R., Doucette-Stamm L., Rubenfield M., Weinstock K., Lee H.M., Dubois J., Rosenthal A., Platzer M., Nyakatura G., Taudien S., Rump A., Yang H., Yu J., Wang J., Huang G., Gu J., Hood L., Rowen L., Madan A., Qin S., Davis R.W., Federspiel N.A., Abola A.P., Proctor M.J., Myers R.M., Schmutz J., Dickson M., Grimwood J., Cox D.R., Olson M.V., Kaul R., Raymond C., Shimizu N., Kawasaki K., Minoshima S., Evans G.A., Athanasiou M., Schultz R., Roe B.A., Chen F., Pan H., Ramser J., Lehrach H., Reinhardt R., McCombie W.R., de la Bastide M., Dedhia N., Blocker H., Hornischer K., Nordsiek G., Agarwala R., Aravind L., Bailey J.A., Bateman A., Batzoglou S., Birney E., Bork P., Brown D.G., Burge C.B., Cerutti L., Chen H.C., Church D., Clamp M., Copley R.R., Doerks T., Eddy S.R., Eichler E.E., Furey T.S., Galagan J., Gilbert J.G., Harmon C., Hayashizaki Y., Haussler D., Hermjakob H. Initial sequencing and analysis of the human genome. Nature. 2001;409:860–921. doi: 10.1038/35057062. [DOI] [PubMed] [Google Scholar]
  38. Lauritsen A., Dahl B.M., Dahl O., Vester B., Wengel J. Methylphosphonate LNA: a locked nucleic acid with a methylphosphonate linkage. Bioorg Med Chem Lett. 2003;13:253–256. doi: 10.1016/S0960-894X(02)00882-X. [DOI] [PubMed] [Google Scholar]
  39. Lee R.C., Ambros V. An extensive class of small RNAs in Caenorhabditis elegans. Science. 2001;294:862–864. doi: 10.1126/science.1065329. [DOI] [PubMed] [Google Scholar]
  40. Lipardi C., Wei Q., Paterson B.M. RNAi as random degradative PCR: siRNA primers convert mRNA into dsRNAs that are degraded to generate new siRNAs. Cell. 2001;107:297–307. doi: 10.1016/S0092-8674(01)00537-2. [DOI] [PubMed] [Google Scholar]
  41. Liss B. Improved quantitative real-time RT-PCR for expression profiling of individual cells. Nucleic Acids Res. 2002;30(17):e89. doi: 10.1093/nar/gnf088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Morita K., Hasegawa C., Kaneko M., Tsutsumi S., Sone J., Ishikawa T., Imanishi T., Koizumi M. 2′-O,4′-C-Ethylene-bridged nucleic acids (ENA): highly nuclease-resistant and thermodynamically stable oligonucleotides for antisense drug. Bioorg Med Chem Lett. 2002;12:73–76. doi: 10.1016/S0960-894X(01)00683-7. [DOI] [PubMed] [Google Scholar]
  43. Mouritzen P., Nielsen P.S., Jacobsen N., Noerholm M., Lomholt C., Pfundheller H.M., Ramsing N.B., Kauppinen S., Tolstrup N. The ProbeLibrary—expression profiling 99% of all human genes using only 90 dual-labeled real-time PCR probes. Biotechniques. 2004;37:492–495. [Google Scholar]
  44. Nielsen K.E., Rasmussen J., Kumar R., Wengel J., Jacobsen J.P., Petersen M. NMR studies of fully modified Locked nucleic acid (LNA) hybrids: solution structure of an LNA:RNA hybrid and characterization of an LNA:DNA hybrid. Bioconjug Chem. 2004;15:449–457. doi: 10.1021/bc034145h. [DOI] [PubMed] [Google Scholar]
  45. Obika S., Nanbu D., Hari Y., Morio K.-I., In Y., Ishida T., Imanishi T. Synthesis of 2′-O,4′-C-methyleneuridine and-cytidine novel bicyclic nucleosides having a fixed C3′-endo sugar puckering. Tetrahedron Lett. 1997;38:8735–8738. doi: 10.1016/S0040-4039(97)10322-7. [DOI] [Google Scholar]
  46. Obika S., Nanbu D., Hari Y., Andoh J.-I., Morio K.-I., Doi T., Imanishi T. Stability and structural features of the duplexes containing nucleoside analogues with a fixed N-type conformation, 2′-O,4′-C-methyleneribonucleosides. Tetrahedron Lett. 1998;39:5401–5404. doi: 10.1016/S0040-4039(98)01084-3. [DOI] [Google Scholar]
  47. Obika S., Hemamayi R., Masuda T., Sugimoto T., Nakagawa S., Mayumi T., Imanishi T. Inhibition of ICAM-1 gene expression by antisense 2′,4′-BNA oligonucleotides. Nucleic Acids Res Suppl. 2001;1:145–146. doi: 10.1093/nass/1.1.145. [DOI] [PubMed] [Google Scholar]
  48. Petersen M., Wengel J. LNA: a versatile tool for therapeutics and genomics. Trends Biotechnol. 2003;21:74–81. doi: 10.1016/S0167-7799(02)00038-0. [DOI] [PubMed] [Google Scholar]
  49. Petersen M., Bondensgaard K., Wengel J., Petersen J.P. Locked nucleic acid (LNA) recognition of RNA: NMR solution structures of LNA:RNA hybrids. J Am Chem Soc. 2002;124:5974–5982. doi: 10.1021/ja012288d. [DOI] [PubMed] [Google Scholar]
  50. Pfundheller H.M., Sorensen A.M., Lomholt C., Johansen A.M., Koch T., Wengel J. Locked nucleic acid synthesis. Methods Mol Biol. 2005;288:127–146. doi: 10.1385/1-59259-823-4:127. [DOI] [PubMed] [Google Scholar]
  51. Reinhart B.J., Slack F.J., Basson M., Pasquinelli A.E., Bettinger J.C., Rougvie A.E., Horvitz H.R., Ruvkun G. The 21 nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature. 2000;403:901–906. doi: 10.1038/35002607. [DOI] [PubMed] [Google Scholar]
  52. Sachidanandam R., Weissman D., Schmidt S.C., Kakol J.M., Stein L.D., Marth G., Sherry S., Mullikin J.C., Mortimore B.J., Willey D.L., Hunt S.E., Cole C.G., Coggill P.C., Rice C.M., Ning Z., Rogers J., Bentley D.R., Kwok P.Y., Mardis E.R., Yeh R.T., Schultz B., Cook L., Davenport R., Dante M., Fulton L., Hillier L., Waterston R.H., McPherson J.D., Gilman B., Schaffner S., Van Etten W.J., Reich D., Higgins J., Daly M.J., Blumenstiel B., Baldwin J., Stange-Thomann N., Zody M.C., Linton L., Lander E.S., Altshuler D., et al. A map of human genome sequence variation containing 1.42 million single nucleotide polymorphisms. Nature. 2001;409:928–933. doi: 10.1038/35057149. [DOI] [PubMed] [Google Scholar]
  53. Santoro S.W., Joyce G.F. A general purpose RNA-cleaving DNA enzyme. Proc Natl Acad Sci USA. 1997;94:4262–4266. doi: 10.1073/pnas.94.9.4262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Schubert S., Gul D.C., Grunert H.P., Zeichhardt H., Erdmann V.A., Kurreck J. RNA cleaving “10–23”DNAzymeswith enhanced stability and activity. Nucleic Acids Res. 2003;31:5982–599. doi: 10.1093/nar/gkg791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Schubert S., Fürste J.P., Werk D., Grunert H.-P., Zeichhardt H., Erdmann V.A., Kurreck J. Gaining target access for deoxyribozymes. J Mol Biol. 2004;339:355–363. doi: 10.1016/j.jmb.2004.03.064. [DOI] [PubMed] [Google Scholar]
  56. Schwarz D.S., Hutvágner G., Du T., Xu Z., Aronin N., Zamore P.D. Asymmetry in the assembly of the RNAi enzyme complex. Cell. 2003;115:199–208. doi: 10.1016/S0092-8674(03)00759-1. [DOI] [PubMed] [Google Scholar]
  57. Singh S.K., Nielsen P., Koshkin A.A., Wengel J. LNA (locked nucleic acids): synthesis and high-affinity nucleic acid recognition. Chem Commun. 1998;4:455–456. [Google Scholar]
  58. Sørensen M.D., Kvaernø L., Bryld T., Håkansson A.E., Verbeure B., Gaubert G., Herdewijn P., Wengel J. α-L-ribo-configured locked nucleic acid (α-L-LNA): synthesis and properties. J Am Chem Soc. 2002;124:2164–2. doi: 10.1021/ja0168763. [DOI] [PubMed] [Google Scholar]
  59. Tolstrup N., Nielsen P.S., Kolberg J.G., Frankel A.M., Vissing H., Kauppinen S. OligoDesign: optimal design of LNA (locked nucleic acid) oligonucleotide capture probes for gene expression profiling. Nucleic Acids Res. 2003;31:3758–3762. doi: 10.1093/nar/gkg580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Valoczi A., Hornyik C., Varga N., Burgyan J., Kauppinen S., Havelda Z. Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. Nucleic Acids Res. 2004;32:e175. doi: 10.1093/nar/gnh171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Venter J.C., Adams M.D., Myers E.W., Li P.W., Mural R.J., Sutton G.G., Smith H.O., Yandell M., Evans C.A., Holt R.A., Gocayne J.D., Amanatides P., Ballew R.M., Huson D.H., Wortman J.R., Zhang Q., Kodira C.D., Zheng X.H., Chen L., Skupski M., Subramanian G., Thomas P.D., Zhang J., Gabor Miklos G.L., Nelson C., Broder S., Clark A.G., Nadeau J., McKusick V.A., Zinder N., Levine A.J., Roberts R.J., Simon M., Slayman C., Hunkapiller M., Bolanos R., Delcher A., Dew I., Fasulo D., Flanigan M., Florea L., Halpern A., Hannenhalli S., Kravitz S., Levy S., Mobarry C., Reinert K., Remington K., Abu-Threideh J., Beasley E., Biddick K., Bonazzi V., Brandon R., Cargill M., Chandramouliswaran I., Charlab R., Chaturvedi K., Deng Z., Di Francesco V., Dunn P., Eilbeck K., Evangelista C., Gabrielian A.E., Gan W., Ge W., Gong F., Gu Z., Guan P., Heiman T.J., Higgins M.E., Ji R.R., Ke Z., Ketchum K.A., Lai Z., Lei Y., Li Z., Li J., Liang Y., Lin X., Lu F., Merkulov G.V., Milshina N., Moore H.M., Naik A.K., Narayan V.A., Neelam B., Nusskern D., Rusch D.B., Salzberg S., Shao W., Shue B., Sun J., Wang Z., Wang A., Wang X., Wang J., Wei M., Wides R., Xiao C., Yan C., Yao A., Ye J., Zhan M., Zhang W., Zhang H., Zhao Q., Zheng L., Zhong F., Zhong W., Zhu S., Zhao S., Gilbert D., Baumhueter S., Spier G., Carter C., Cravchik A., Woodage T., Ali F., An H., Awe A., Baldwin D., Baden H., Barnstead M., Barrow I., Beeson K., Busam D., Carver A., Center A., Cheng M.L., Curry L., Danaher S., Davenport L., Desilets R., Dietz S., Dodson K., Doup L., Ferriera S., Garg N., Gluecksmann A., Hart B., Haynes J., Haynes C., Heiner C., Hladun S., Hostin D., Houck J., Howland T., Ibegwam C., Johnson J., Kalush F., Kline L., Koduru S., Love A., Mann F., May D., McCawley S., McIntosh T., McMullen I., Moy M., Moy L., Murphy B., Nelson K., Pfannkoch C., Pratts E., Puri V., Qureshi H., Reardon M., Rodriguez R., Rogers Y.H., Romblad D., Ruhfel B., Scott R., Sitter C., Smallwood M., Stewart E., Strong R., Suh E., Thomas R., Tint N.N., Tse S., Vech C., Wang G., Wetter J., Williams S., Williams M., Windsor S., Winn-Deen E., Wolfe K., Zaveri J., Zaveri K., Abril J.F., Guigo R., Campbell M.J., Sjolander K.V., Karlak B., Kejariwal A., Mi H., Lazareva B., Hatton T., Narechania A., Diemer K., Muruganujan A., Guo N., Sato S., Bafna V., Istrail S., Lippert R., Schwartz R., Walenz B., Yooseph S., Allen D. The sequence of the human genome. Science. 2001;291:1304–1351. doi: 10.1126/science.1058040. [DOI] [PubMed] [Google Scholar]
  62. Vester B., Wengel J. LNA (locked nucleic acid): high affinity targeting of complementary RNA and DNA. Biochemistry. 2004;43:13233–13241. doi: 10.1021/bi0485732. [DOI] [PubMed] [Google Scholar]
  63. Vester B., Lundberg L., Sørensen M.D., Babu B.R., Douthwaite S., Wengel J. LNAzymes: incorporation of LNA-type monomers into DNAzymes markedly increases RNA cleavage. J Am Chem Soc. 2002;124:13682–13683. doi: 10.1021/ja0276220. [DOI] [PubMed] [Google Scholar]
  64. Wahlestedt C., Salmi P., Good L., Kela J., Johnsson T., Hokfelt T., Broberger C., Porreca F., Lai J., Ren K., Ossipov M., Koshkin A., Jakobsen N., Skouv J., Oerum H., Jacobsen M.H., Wengel J. Potent and nontoxic antisense oligonucleotides containing locked nucleic acids. Proc Natl Acad Sci USA. 2000;97:5633–5638. doi: 10.1073/pnas.97.10.5633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Wengel J. Synthesis of 3′-C-and 4′-C-branched oligonucleotides and the development of locked nucleic acid (LNA) Acc Chem Res. 1999;32:301–310. doi: 10.1021/ar980051p. [DOI] [Google Scholar]
  66. Wienholds E., Kloosterman W.P., Miska E., Alvarez-Saavedra E., Berezikov E., de Bruijn E., Horvitz H.R., Kauppinen S., Plasterk R.H.A. MicroRNA expression in zebrafish embryonic development. Science. 2005;309:310–311. doi: 10.1126/science.1114519. [DOI] [PubMed] [Google Scholar]
  67. Wong G.K., Passey D.G., Yu J. Most of the human genome is transcribed. Genome Res. 2001;11:1975–1977. doi: 10.1101/gr.202401. [DOI] [PubMed] [Google Scholar]

Articles from RNA Towards Medicine are provided here courtesy of Nature Publishing Group

RESOURCES