Peptide Guided Gene Delivery

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Omai's picture
Peptide Guided Gene Delivery

Check out this great paper on using peptides to guide gene delivery. Most in vivo gene delivery systems use viruses to deliver genetic cargo to a particular cell. A peptide delivery system would have to meet certain goals including protecting the DNA, targeting specific cell surface receptors, disrupting the endosomal membrane, and delivering the cargo DNA to the nucleus. This paper has a table listing several peptides that can meet these four goals. Here is a copy of figure 1 that outlines these conditions:

The paper goes into great detail about using peptides for gene delivery, specifically talking about receptor specificity, endosomal escape, and nuclear localization. I had never even heard of using a protcetive peptide for gen therapy.
Very cool. Give the article a read if you have the chance.

Jon Moulton
Jon Moulton's picture
A variation on this is used

A variation on this is used for delivery of Morpholino antisense.  In this application, peptides are covalently linked to the oligos (necessary because the Morpholinos have no charge, so they won't form electrostatic complexes).  The oligos are taken up systemically and are active in the nucleus and cytosol.  Peptide conjugates of phosphorodiamidate Morpholino oligos are called PPMO.   I keep track of this literature -- the inventor, Hong Moulton (Moulton HM), is my wife.  Here are some references for the Morpholino-peptide conjugate system.
Moulton JD, Jiang S. Gene Knockdowns in Adult Animals: PPMOs and Vivo-Morpholinos. Molecules. 2009 Mar 25;14(3):1304-23.
Svasti S, Suwanmanee T, Fucharoen S, Moulton HM, Nelson MH, Maeda N, Smithies O, Kole R.  RNA repair restores hemoglobin expression in IVS2-654 thalassemic mice.  Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1205-10. Epub 2009 Jan 21.
Swenson DL, Warfield KL, Warren TK, Lovejoy C, Hassinger JN, Ruthel G, Blouch RE, Moulton HM, Weller DD, Iversen PL, Bavari S.  Chemical modifications of antisense morpholino oligomers enhance their efficacy against ebolavirus infection.  Antimicrob Agents Chemother. 2009 Feb 17. [Epub ahead of print]
Zhang B, Dong H, Stein DA, Shi PY.  Co-selection of West Nile virus nucleotides that confer resistance to an antisense oligomer while maintaining long-distance RNA/RNA base pairings.  Virology. 2008 Oct 6. [Epub ahead of print]
Abes R, Moulton HM, Clair P, Yang ST, Abes S, Melikov K, Prevot P, Youngblood DS, Iversen PL, Chernomordik LV, Lebleu B.  Delivery of steric block morpholino oligomers by (R-X-R)4 peptides: structure-activity studies.  Nucleic Acids Res. 2008 Sep 16. [Epub ahead of print]
Bok K, Cavanaugh VJ, Matson DO, González-Molleda L, Chang KO, Zintz C, Smith AW, Iversen P, Green KY, Campbell AE.  Inhibition of norovirus replication by morpholino oligomers targeting the 5'-end of the genome.  Virology. 2008 Sep 8. [Epub ahead of print]
Yin H, Moulton HM, Seow Y, Boyd C, Boutilier J, Iverson P, Wood MJ.  Cell-penetrating peptide-conjugated antisense oligonucleotides restore systemic muscle and cardiac dystrophin expression and function.  Hum Mol Genet. 2008 Sep 10. [Epub ahead of print]
Wu B, Moulton HM, Iversen PL, Jiang J, Li J, Li J, Spurney CF, Sali A, Guerron AD, Nagaraju K, Doran T, Lu P, Xiao X, Lu QL.  Effective rescue of dystrophin improves cardiac function in dystrophin-deficient mice by a modified morpholino oligomer.  Proc Natl Acad Sci U S A. 2008 Sep 19. [Epub ahead of print]
Jearawiriyapaisarn N, Moulton HM, Buckley B, Roberts J, Sazani P, Fucharoen S, Iversen PL, Kole R.  Sustained Dystrophin Expression Induced by Peptide-conjugated Morpholino Oligomers in the Muscles of mdx Mice.  Mol Ther. 2008 Sep;16(9):1624-9. Epub 2008 Jun 10.
Paessler S, Rijnbrand R, Stein DA, Ni H, Yun NE, Dziuba N, Borisevich V, Seregin A, Ma Y, Blouch R, Iversen PL, Zacks MA.  Inhibition of alphavirus infection in cell culture and in mice with antisense morpholino oligomers.  Virology. 2008 Jul 5;376(2):357-70. Epub 2008 May 12.
Abes R, Arzumanov A, Moulton H, Abes S, Ivanova G, Gait MJ, Iversen P, Lebleu B.  Arginine-rich cell penetrating peptides: Design, structure-activity, and applications to alter pre-mRNA splicing by steric-block oligonucleotides.  J Pept Sci. 2008 Apr;14(4):455-60.
Lupfer C, Stein DA, Mourich DV, Tepper SE, Iversen PL, Pastey M.  Inhibition of influenza A H3N8 virus infections in mice by morpholino oligomers.  Arch Virol. 2008 May;153(5):929-37. Epub 2008 Mar 28.
Patel D, Opriessnig T, Stein DA, Halbur PG, Meng XJ, Iversen PL, Zhang YJ.  Peptide-conjugated morpholino oligomers inhibit porcine reproductive and respiratory syndrome virus replication.  Antiviral Res. 2007 Oct 4; [Epub ahead of print]
Lebleu B, Moulton HM, Abes R, Ivanova GD, Abes S, Stein DA, Iversen PL, Arzumanov AA, Gait MJ.  Cell penetrating peptide conjugates of steric block oligonucleotides.  Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):517-29. Epub 2007 Oct 22.
Lai SH, Stein DA, Guerrero-Plata A, Liao SL, Ivanciuc T, Hong C, Iversen PL, Casola A, Garofalo RP.  Inhibition of Respiratory Syncytial Virus Infections With Morpholino Oligomers in Cell Cultures and in Mice.  Mol Ther. 2008 Apr 29; [Epub ahead of print]
Zhang YJ, Bonaparte RS, Patel D, Stein DA, Iversen PL.  Blockade of viral interleukin-6 expression of Kaposi's sarcoma-associated herpesvirus.  Mol Cancer Ther. 2008 Mar;7(3):712-20.
Stone JK, Rijnbrand R, Stein DA, Ma Y, Yang Y, Iversen PL, Andino R.  A morpholino oligomer targeting highly conserved internal ribosome entry site sequence is able to inhibit multiple species of picornavirus.  Antimicrob Agents Chemother. 2008 Jun;52(6):1970-81. Epub 2008 Mar 17.
Gabriel G, Nordmann A, Stein DA, Iversen PL, Klenk HD.  Morpholino oligomers targeting the PB1 and NP genes enhance the survival of mice infected with highly pathogenic influenza A H7N7 virus.  J Gen Virol. 2008 Apr;89(Pt 4):939-48.
Vagnozzi A, Stein DA, Iversen PL, Rieder E.  Inhibition of foot-and-mouth disease virus in cell cultures with antisense Morpholino oligomers.  J Virol. 2007 Aug 29; [Epub ahead of print]
Marshall NB, Oda SK, London CA, Moulton HM, Iversen PL, Kerkvliet NI, Mourich DV.  Arginine-rich cell-penetrating peptides facilitate delivery of antisense oligomers into murine leukocytes and alter pre-mRNA splicing.  J Immunol Methods. 2007 Jul 26; [Epub ahead of print]
Wu RP, Youngblood DS, Hassinger JN, Lovejoy CE, Nelson MH, Iversen PL, Moulton HM.  Cell-penetrating peptides as transporters for morpholino oligomers: effects of amino acid composition on intracellular delivery and cytotoxicity.  Nucleic Acids Res. 2007 Aug 1; [Epub ahead of print]
Moulton HM, Fletcher S, Neuman BW, McClorey G, Stein DA, Abes S, Wilton SD, Buchmeier MJ, Lebleu B, Iversen PL.  Cell-penetrating peptide-morpholino conjugates alter pre-mRNA splicing of DMD (Duchenne muscular dystrophy) and inhibit murine coronavirus replication in vivo.  Biochem Soc Trans. 2007 Aug;35(Pt 4):826-8.
Amantana A, Moulton HM, Cate ML, Reddy MT, Whitehead T, Hassinger JN, Youngblood DS, Iversen PL.  Pharmacokinetics, Biodistribution, Stability and Toxicity of a Cell-Penetrating Peptide-Morpholino Oligomer Conjugate.  Bioconjug Chem. 2007 Jun 21; [Epub ahead of print]
Fletcher S, Honeyman K, Fall AM, Harding PL, Johnsen RD, Steinhaus JP, Moulton HM, Iversen PL, Wilton SD.  Morpholino Oligomer-Mediated Exon Skipping Averts the Onset of Dystrophic Pathology in the mdx Mouse.  Mol Ther. 2007 Jun 19; [Epub ahead of print]
Deas TS, Bennett CJ, Jones SA, Tilgner M, Ren P, Behr MJ, Stein DA, Iversen PL, Kramer LD, Bernard KA, Shi PY.  In vitro resistance selection and in vivo efficacy of morpholino oligomers against West Nile virus.  Antimicrob Agents Chemother. 2007 May 7; [Epub ahead of print] {in-vivo mouse data}
Burrer R, Neuman BW, Ting JPC, Stein DA, Moulton HM, Iversen PL, Kuhn P, Buchmeier MJ.  Antiviral effects of antisense morpholino oligomers in murine coronavirus infection models.  J. Virol. 2007 Epub 2007 Mar 5.{in-vivo mouse data}
Yuan J, Stein DA, Lim T, Qiu D, Coughlin S, Liu Z, Wang Y, Bouch R, Moulton HM, Iversen PL, Yang D.   Inhibition of Coxsackievirus B3 in Cell-cultures and in Mice by Peptide-Conjugated Morpholino Oligomers Targeting the IRES.  J Virol. 2006 Sep 20; [Epub ahead of print]
Abes S, Moulton HM, Clair P, Prevot P, Youngblood DS, Wu RP, Iversen PL, Lebleu B.  Vectorization of morpholino oligomers by the (R-Ahx-R)(4) peptide allows efficient splicing correction in the absence of endosomolytic agents.  J Control Release. 2006 Sep 30; [Epub ahead of print] {nice mechanism study, also a good place to start on how to conjugate}
Youngblood DS, Hatlevig SA, Hassinger JN, Iversen PL, Moulton HM.  Stability of cell-penetrating Peptide-morpholino oligomer conjugates in human serum and in cells.  Bioconjug Chem. 2007 Jan-Feb;18(1):50-60.
McClorey G, Fall AM, Moulton HM, Iversen PL, Rasko JE, Ryan M, Fletcher S, Wilton SD.  Induced dystrophin exon skipping in human muscle explants.  Neuromuscul Disord. 2006 Oct;16(9-10):583-90. Epub 2006 Aug 21 {application paper, little background on the conjugates}
McClorey G, Moulton HM, Iversen PL, Fletcher S, Wilton SD.  Antisense oligonucleotide-induced exon skipping restores dystrophin expression in vitro in a canine model of DMD.  Gene Ther. 2006 Oct;13(19):1373-81. Epub 2006 May 25 {application paper, little background on the conjugates}
Deas TS, Binduga-Gajewska I, Tilgner M, Ren P, Stein DA, Moulton HM, Iversen PL, Kauffman EB, Kramer LD, Shi PY.  Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication.  J Virol. 2005 Apr;79(8):4599-609 {application paper, little background on the conjugates}
Nelson MH, Stein DA, Kroeker AD, Hatlevig SA, Iversen PL, Moulton HM.  Arginine-rich peptide conjugation to morpholino oligomers: effects on antisense activity and specificity.  Bioconjug Chem. 2005 Jul-Aug;16(4):959-66. {Nice paper specifically on the conjugates}
Moulton HM, Moulton JD.  Arginine-rich cell-penetrating peptides with uncharged antisense oligomers.  Drug Discov Today. 2004 Oct 15;9(20):870. {argument for the effectiveness of the Morpholino-peptide system}
Moulton HM, Nelson MH, Hatlevig SA, Reddy MT, Iversen PL.  Cellular uptake of antisense morpholino oligomers conjugated to arginine-rich peptides.  Bioconjug Chem. 2004 Mar-Apr;15(2):290-9. {Nice paper specifically on the conjugates}
Moulton HM, Moulton JD.  Peptide-assisted delivery of steric-blocking antisense oligomers.  Curr Opin Mol Ther. 2003 Apr;5(2):123-32. {review}
Moulton HM, Hase MC, Smith KM, Iversen PL.  HIV Tat peptide enhances cellular delivery of antisense morpholino oligomers.  Antisense Nucleic Acid Drug Dev. 2003 Feb;13(1):31-43. {early exploration with less-optimal peptide}