Receptor-mediated delivery of siRNAs by tethered nucleic acid base-paired interactions - PubMed (original) (raw)
Receptor-mediated delivery of siRNAs by tethered nucleic acid base-paired interactions
Kexiong Zhang et al. RNA. 2008 Mar.
Abstract
We report a new strategy for cell-type-specific delivery of functional siRNAs into cells. The method involves the noncovalent attachment of siRNAs to ligand-conjugated oligodeoxynucleotides via nucleic acid base-paired interactions. The resulting complexes can be directly applied to cells, leading to specific cellular uptake and gene silencing. The method is simple, economical, and can be easily adapted for other cell surface receptors. Here we show the application of this method for the delivery of siRNAs to folate receptor-expressing cells.
Figures
FIGURE 1.
Schematic illustration of the tethering strategy and proposed mechanism of action. The siRNAs specific for the human αV (αV) integrin and survivin (Sur) mRNAs, and a nonsilencing control siRNA used in the studies are shown on the left. The 5′-end of the tethering ODN is covalently conjugated to folate (blue dot). The antisense RNA strands complementary to the target mRNAs are in green. The 3′-ends of the sense RNA strands (red) are extended to form base-paired interactions with the ODN. Following entry into the cell via FR-mediated endocytosis, the siRNA complexes (F-ODN:siRNA) are processed and the released siRNAs incorporated into RNA-induced silencing complex (RISC).
FIGURE 2.
FR expression. (A–C) The representative results of immunofluorescence. The anti-FR staining is in red and the nuclear DAPI staining is in blue. (D–F) The representative results of the folate-delivery assay. The green dots indicate cytoplasmic accumulation of folate-oregon green 488, while the nuclear DAPI staining is in blue.
FIGURE 3.
Cell-type specific gene silencing by tethered siRNAs. HUVECs (A,C,D) and MDA-MB-435S (B) cells were incubated with buffer alone, or with 300 nM of the various siRNAs in the presence of 10% fetal bovine serum. Four hours later, the media were removed and cells were incubated in fresh media containing no siRNAs for an additional 22 h. In C, last column, HUVECs were transfected with poly (I:C) using Lipofectamine 2000. In A–C, total cellular RNAs were extracted and the indicated mRNA levels measured by qRT_–_PCR. Each bar represents mean ± sem (n = 3–6). In D, cell viability and caspase activity of HUVECs were measured following the various treatments above, and the results of an average of two independent experiments were plotted. (Bfr) Buffer alone; (F-ODN:siRNA) siRNA tethered by the folate-conjugated ODN; (F-ODN:siRNA+free folate) F-ODN:siRNA in the presence of 200x molar excess free folate; (ODN:siRNA) siRNA tethered by the same ODN but without folate conjugated; Poly (I:C) cells transfected with poly (I:C).
FIGURE 4.
Specific gene silencing by tethered siRNAs in KB cells. KB cells were incubated with buffer alone, or with 300 nM of the various siRNAs in the presence of 10% fetal bovine serum. Four hours later, the media were removed and cells incubated in fresh media containing no siRNAs for an additional 22 h. In B, last column, KB cells were transfected with poly (I:C) using Lipofectamine 2000. Total cellular RNAs were extracted and the indicated mRNA levels measured by qRT–PCR. Each bar represents mean±sem (n = 3–6). Other labels are the same as in Fig. 3.
Similar articles
- Efficient intracellular delivery of oligonucleotides formulated in folate receptor-targeted lipid vesicles.
Zhou W, Yuan X, Wilson A, Yang L, Mokotoff M, Pitt B, Li S. Zhou W, et al. Bioconjug Chem. 2002 Nov-Dec;13(6):1220-5. doi: 10.1021/bc025569z. Bioconjug Chem. 2002. PMID: 12440856 - Silencing of survivin gene by small interfering RNAs produces supra-additive growth suppression in combination with 17-allylamino-17-demethoxygeldanamycin in human prostate cancer cells.
Paduano F, Villa R, Pennati M, Folini M, Binda M, Daidone MG, Zaffaroni N. Paduano F, et al. Mol Cancer Ther. 2006 Jan;5(1):179-86. doi: 10.1158/1535-7163.MCT-05-0132. Mol Cancer Ther. 2006. PMID: 16432177 - Silencing stathmin gene expression by survivin promoter-driven siRNA vector to reverse malignant phenotype of tumor cells.
Zhang HZ, Wang Y, Gao P, Lin F, Liu L, Yu B, Ren JH, Zhao H, Wang R. Zhang HZ, et al. Cancer Biol Ther. 2006 Nov;5(11):1457-61. doi: 10.4161/cbt.5.11.3272. Epub 2006 Nov 13. Cancer Biol Ther. 2006. PMID: 17012855 - Knockdown of survivin expression by siRNAs enhances chemosensitivity of prostate cancer cells and attenuates its tumorigenicity.
Shen J, Liu J, Long Y, Miao Y, Su M, Zhang Q, Han H, Hao X. Shen J, et al. Acta Biochim Biophys Sin (Shanghai). 2009 Mar;41(3):223-30. doi: 10.1093/abbs/gmp005. Acta Biochim Biophys Sin (Shanghai). 2009. PMID: 19280061 - Folate-mediated targeting of antineoplastic drugs, imaging agents, and nucleic acids to cancer cells.
Wang S, Low PS. Wang S, et al. J Control Release. 1998 Apr 30;53(1-3):39-48. doi: 10.1016/s0168-3659(97)00236-8. J Control Release. 1998. PMID: 9741912 Review.
Cited by
- Ligand-mediated delivery of RNAi-based therapeutics for the treatment of oncological diseases.
Abdelaal AM, Kasinski AL. Abdelaal AM, et al. NAR Cancer. 2021 Jul 20;3(3):zcab030. doi: 10.1093/narcan/zcab030. eCollection 2021 Sep. NAR Cancer. 2021. PMID: 34316717 Free PMC article. - FolamiRs: Ligand-targeted, vehicle-free delivery of microRNAs for the treatment of cancer.
Orellana EA, Tenneti S, Rangasamy L, Lyle LT, Low PS, Kasinski AL. Orellana EA, et al. Sci Transl Med. 2017 Aug 2;9(401):eaam9327. doi: 10.1126/scitranslmed.aam9327. Sci Transl Med. 2017. PMID: 28768807 Free PMC article. - New type of BACE1 siRNA delivery to cells.
Jabłkowski M, Szemraj M, Oszajca K, Janiszewska G, Bartkowiak J, Szemraj J. Jabłkowski M, et al. Med Sci Monit. 2014 Dec 10;20:2598-606. doi: 10.12659/MSM.891219. Med Sci Monit. 2014. PMID: 25491230 Free PMC article. - Site-specific antibody-polymer conjugates for siRNA delivery.
Lu H, Wang D, Kazane S, Javahishvili T, Tian F, Song F, Sellers A, Barnett B, Schultz PG. Lu H, et al. J Am Chem Soc. 2013 Sep 18;135(37):13885-91. doi: 10.1021/ja4059525. Epub 2013 Sep 5. J Am Chem Soc. 2013. PMID: 23924037 Free PMC article. - Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
Jung HJ, Lim JS, Choi HJ, Lee MS, Kim JH, Kim SY, Kim S, Kim E, Kwon TH. Jung HJ, et al. PLoS One. 2012;7(6):e40010. doi: 10.1371/journal.pone.0040010. Epub 2012 Jun 28. PLoS One. 2012. PMID: 22761946 Free PMC article.
References
- Aronov, O., Horowitz, A.T., Gabizon, A., Gibson, D. Folate-targeted PEG as a potential carrier for carboplatin analogs. Synthesis and in vitro studies. Bioconjug. Chem. 2003;14:563–574. - PubMed
- Cao, Q., Cai, W., Li, T., Yang, Y., Chen, K., Xing, L., Chen, X. Combination of integrin siRNA and irradiation for breast cancer therapy. Biochem. Biophys. Res. Commun. 2006;351:726–732. - PubMed
- Hu-Lieskovan, S., Heidel, J.D., Bartlett, D.W., Davis, M.E., Triche, T.J. Sequence-specific knockdown of EWS-FLI1 by targeted, nonviral delivery of small interfering RNA inhibits tumor growth in a murine model of metastatic Ewing's sarcoma. Cancer Res. 2005;65:8984–8992. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources