title: Activation of different split functionalities on re-association of RNA-DNA hybrids.
abstract: Split-protein systems, an approach that relies on fragmentation of proteins with their further conditional re-association to form functional complexes, are increasingly used for various biomedical applications. This approach offers tight control of protein functions and improved detection sensitivity. Here we report a similar technique based on a pair of RNA-DNA hybrids that can be used generally for triggering different split functionalities. Individually, each hybrid is inactive but when two cognate hybrids re-associate, different functionalities are triggered inside mammalian cells. As a proof of concept, this work mainly focuses on the activation of RNA interference. However, the release of other functionalities (such as resonance energy transfer and RNA aptamer) is also shown. Furthermore, in vivo studies demonstrate a significant uptake of the hybrids by tumours together with specific gene silencing. This split-functionality approach presents a new route in the development of 'smart' nucleic acid-based nanoparticles and switches for various biomedical applications.
authors.affiliation: Center for Cancer Research Nanobiology Program, NCI, Frederick, Maryland 21702, USA.
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Afonin
authors.forename: Mathias
authors.initials: M
authors.lastname: Viard
authors.forename: Angelica N
authors.initials: AN
authors.lastname: Martins
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Lockett
authors.forename: Anna E
authors.initials: AE
authors.lastname: Maciag
authors.forename: Eric O
authors.initials: EO
authors.lastname: Freed
authors.forename: Eliahu
authors.initials: E
authors.lastname: Heldman
authors.forename: Luc
authors.initials: L
authors.lastname: Jaeger
authors.forename: Robert
authors.initials: R
authors.lastname: Blumenthal
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Shapiro
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D018613:Microscopy, Confocal; D009692:Nucleic Acid Heteroduplexes / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
chemical_list: D052157:Aptamers, Nucleotide; D009692:Nucleic Acid Heteroduplexes; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D012313:RNA; D004247:DNA
references.citation: Nat Nanotechnol. 2010 Dec;5(12):833-42
references.pmid: 21102465
references.citation: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10340-4
references.pmid: 7937952
references.citation: J Mol Biol. 2000 Aug 4;301(1):117-28
references.pmid: 10926496
references.citation: EMBO J. 2001 Dec 3;20(23):6877-88
references.pmid: 11726523
references.citation: Nature. 2004 Sep 16;431(7006):371-8
references.pmid: 15372045
references.citation: Proc Natl Acad Sci U S A. 1999 May 25;96(11):6131-6
references.pmid: 10339553
references.citation: Mol Ther Nucleic Acids. 2013 Mar 19;2:e80
references.pmid: 23511334
references.citation: Nat Methods. 2011 Nov 29;8(12):990-2
references.pmid: 22127214
references.citation: PLoS One. 2011;6(5):e20359
references.pmid: 21647381
references.citation: Nature. 2009 Jan 22;457(7228):405-12
references.pmid: 19158786
references.citation: Nat Nanotechnol. 2010 Sep;5(9):676-82
references.pmid: 20802494
references.citation: ACS Nano. 2011 May 24;5(5):3405-18
references.pmid: 21604810
references.citation: RNA. 2003 Sep;9(9):1034-48
references.pmid: 12923253
references.citation: Curr Opin Chem Biol. 2011 Dec;15(6):789-97
references.pmid: 22070901
references.citation: Nature. 2004 Sep 16;431(7006):343-9
references.pmid: 15372041
references.citation: Mol Ther. 2012 Apr;20(4):820-8
references.pmid: 22314289
references.citation: Nat Rev Cancer. 2011 Jan;11(1):59-67
references.pmid: 21160526
references.citation: Nano Lett. 2011 Feb 9;11(2):878-87
references.pmid: 21229999
references.citation: Methods. 2011 Jun;54(2):204-14
references.pmid: 21320601
references.citation: Mol Pharm. 2009 May-Jun;6(3):659-68
references.pmid: 19267452
references.citation: Nucleic Acids Res. 1979 Aug 10;6(11):3543-57
references.pmid: 158748
references.citation: J Comput Chem. 2011 Jan 15;32(1):170-3
references.pmid: 20645303
references.citation: Proc Natl Acad Sci U S A. 2010 Sep 28;107(39):16777-82
references.pmid: 20823260
references.citation: Biophys J. 2003 Jun;84(6):3992-4010
references.pmid: 12770904
references.citation: Nucleic Acids Res. 2005 Jul 26;33(13):4140-56
references.pmid: 16049023
references.citation: Curr Biol. 2003 Jan 8;13(1):41-6
references.pmid: 12526743
references.citation: Biochemistry. 1995 Sep 5;34(35):11211-6
references.pmid: 7545436
references.citation: Nat Protoc. 2011 Dec 01;6(12):2022-34
references.pmid: 22134126
references.citation: Science. 2007 Nov 16;318(5853):1121-5
references.pmid: 18006742
references.citation: Nucleic Acids Res. 2007;35(17):5886-97
references.pmid: 17726057
references.citation: Chembiochem. 2008 Aug 11;9(12):1902-5
references.pmid: 18655086
references.citation: Oncogene. 2003 Oct 20;22(47):7369-75
references.pmid: 14576844
references.citation: EMBO J. 2002 Nov 1;21(21):5875-85
references.pmid: 12411505
references.citation: Curr Opin Struct Biol. 2010 Feb;20(1):90-7
references.pmid: 20053548
references.citation: Nucleic Acids Res. 2012 Mar;40(5):2168-80
references.pmid: 22080507
references.citation: Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
references.pmid: 12824337
references.citation: Nat Nanotechnol. 2011 Sep 11;6(10):658-67
references.pmid: 21909084
references.citation: Nano Lett. 2012 Oct 10;12(10):5192-5
references.pmid: 23016824
references.citation: Nature. 1998 Feb 19;391(6669):806-11
references.pmid: 9486653
references.citation: Nature. 2010 Apr 15;464(7291):1067-70
references.pmid: 20305636
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 19;108(16):6405-8
references.pmid: 21464284
references.citation: Pharm Res. 2011 Dec;28(12):2996-3015
references.pmid: 22033880
references.citation: Nat Rev Mol Cell Biol. 2007 Jan;8(1):23-36
references.pmid: 17183358
references.citation: J Am Chem Soc. 2006 Dec 20;128(50):16131-7
references.pmid: 17165766
references.citation: Antiviral Res. 2011 Oct;92(1):7-14
references.pmid: 21513746
references.citation: Mol Cell. 2000 Nov;6(5):1077-87
references.pmid: 11106747
references.citation: J Am Chem Soc. 2004 Aug 4;126(30):9266-70
references.pmid: 15281816
references.citation: Methods Mol Biol. 2009;485:163-84
references.pmid: 19020825
references.citation: Nat Biotechnol. 2005 Feb;23(2):222-6
references.pmid: 15619617
references.citation: Science. 2004 Dec 17;306(5704):2068-72
references.pmid: 15604402