The translation of interfering RNA to the clinic requires more effective delivery agents to enable safe and efficient delivery. The aim of this work was to create a multi-functional delivery agent using deactivation enhanced ATRP synthesis of poly(dimethylamino)ethyl methacrylate (pDMAEMA)-co-PEGMEA/PEGDA (pD-b-P/DA) with linear pDMAEMA as a macro-initiator. The pD-b-P/DA was characterized for its potential to bind synthetic microRNA mimics to form structures and reacted with antibody-derived fragments (Fabs) using Michael-type addition. Conjugation of antibody fragments was verified using SDS–PAGE. Functional delivery of these interfering RNA complexes was proven using a dual luciferase reporter assay. Functional silencing of a reporter gene was improved by complexation of microRNA mimics with pD-b-P/DA alone and with Fab-decorated pD-b-P/DA. The improved silencing with Fab-decorated pD-b-P/DA was evident at 48 h but disappeared at 96 h. The resultant agent enables complexation of nucleic acid (microRNA mimic) and facile conjugation of antibody fragments via a Michael-type addition. In conclusion, this platform is effective at silencing in this reporter system and has potential as an effective delivery system of interfering RNA.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Shan Z-X, Lin Q-X, Fu Y-H, Deng C-Y, Zhou Z-L, Zhu J-N, et al. Upregulated expression of miR-1/miR-206 in a rat model of myocardial infarction. Biochem Biophys Res Commun. 2009;381(4):597–601.
Trang P, Wiggins JF, Daige CL, Cho C, Omotola M, Brown D, et al. Systemic delivery of tumor suppressor microRNA mimics using a neutral lipid emulsion inhibits lung tumors in mice. Mol Ther. 2011;19(6):1116–22.
van Rooij E, Sutherland LB, Thatcher JE, DiMaio JM, Naseem RH, Marshall WS, et al. Dysregulation of microRNAs after myocardial infarction reveals a role of miR-29 in cardiac fibrosis. Proc Natl Acad Sci U S A. 2008;105(35):13027–32.
Yekta S, I-h S, Bartel DP. MicroRNA-directed cleavage of HOXB8 mRNA. Science. 2004;304(5670):594–6.
Monaghan M, Pandit A. RNA interference therapy via functionalized scaffolds. Adv Drug Deliv Rev. 2011;63(4–5):197–208.
Manjunath N, Wu H, Subramanya S, Shankar P. Lentiviral delivery of short hairpin RNAs. Adv Drug Deliv Rev. 2009;61(9):732–45.
Tomlinson E, Rolland AP. Controllable gene therapy pharmaceutics of non-viral gene delivery systems. J Control Release. 1996;39(2–3):357–72.
Rahbek UL, Howard KA, Oupicky D, Manickam DS, Dong M, Nielsen AF, et al. Intracellular siRNA and precursor miRNA trafficking using bioresponsive copolypeptides. J Gene Med. 2008;10(1):81–93.
Xu D, Takeshita F, Hino Y, Fukunaga S, Kudo Y, Tamaki A, et al. miR-22 represses cancer progression by inducing cellular senescence. J Cell Biol. 2011;193(2):409–24.
Pramanik D, Campbell NR, Karikari C, Chivukula R, Kent OA, Mendell JT, et al. Restitution of tumor suppressor microRNAs using a systemic nanovector inhibits pancreatic cancer growth in mice. Mol Cancer Ther. 2011;10(8):1470–80.
Huang Y-H, Bao Y, Peng W, Goldberg M, Love K, Bumcrot DA, et al. Claudin-3 gene silencing with siRNA suppresses ovarian tumor growth and metastasis. Proc Natl Acad Sci U S A. 2009;106(9):3426–30.
Davis ME, Zuckerman JE, Choi CHJ, Seligson D, Tolcher A, Alabi CA, et al. Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature. 2010;464(7291):1067–70.
Kim SH, Jeong JH, Lee SH, Kim SW, Park TG. Local and systemic delivery of VEGF siRNA using polyelectrolyte complex micelles for effective treatment of cancer. J Control Release. 2008;129(2):107–16.
Nelson AL, Dhimolea E, Reichert JM. Development trends for human monoclonal antibody therapeutics. Nat Rev Drug Discov. 2010;9(10):767–74.
Heggli M, Tirelli N, Zisch A, Hubbell JA. Michael-type addition as a tool for surface functionalization. Bioconjug Chem. 2003;14(5):967–73.
Zheng Y, Cao H, Newland B, Dong Y, Pandit A, Wang W. 3D single cyclized polymer chain structure from controlled polymerization of multi-vinyl monomers: beyond Flory–Stockmayer theory. J Am Chem Soc. 2011;133(33):13130–7.
Newland B, Zheng Y, Jin Y, Abu-Rub M, Cao H, Wang W, et al. Single cyclized molecule versus single branched molecule: a simple and efficient 3D “knot” polymer structure for nonviral gene delivery. J Am Chem Soc. 2012;134(10):4782–9.
Newland B, Tai H, Zheng Y, Velasco D, Di Luca A, Howdle SM, et al. A highly effective gene delivery vector—hyperbranched poly(2-(dimethylamino)ethyl methacrylate) from in situ deactivation enhanced ATRP. Chem Commun. 2010;46(26):4698–700.
Lu B, Mahmud H, Maass AH, Yu B, van Gilst WH, de Boer RA, et al. The Plk1 inhibitor BI 2536 temporarily arrests primary cardiac fibroblasts in mitosis and generates aneuploidy. PLoS One. 2010;5(9):e12963.
O’Dwyer R, Razzaque R, Hu X, Hollingshead S, Wall J. Engineering of cysteine residues leads to improved production of a human dipeptidase enzyme in E. coli. Appl Biochem Biotechnol. 2009;159(1):178–90.
Acknowledgement is given to Science Foundation Ireland, grant no. 07/SRC/B1163, and the College of Engineering and Informatics, National University of Ireland, Galway. Acknowledgement is also due to Dr. Yu Zheng and Ms. Asha Matthew for technical advice.
Electronic supplementary material
Below is the link to the electronic supplementary material.
(DOCX 4356 kb)
About this article
Cite this article
Monaghan, M., Greiser, U., Cao, H. et al. An antibody fragment functionalized dendritic PEGylated poly(2-(dimethylamino)ethyl diacrylate) as a vehicle of exogenous microRNA. Drug Deliv. and Transl. Res. 2, 406–414 (2012). https://doi.org/10.1007/s13346-012-0097-8
- RNA interference
- Dendritic polymers
- Antibody conjugation
- Deactivation enhanced ATRP
- Gene therapy