Mechanisms and Barriers to RNAi Delivery

Zhou, Jiehua; Rossi, John J.

doi:10.1002/9781118610749.ch1

Cited by 5 publications

(2 citation statements)

References 96 publications

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“…6,7 , are subject to rapid renal clearance and degradation by endogenous RNases, and can be recognized by the innate immune system. [8][9][10] To overcome these limitations, recent studies have focused on the combination of a target-speci c antibody and a cell-penetrating peptide with siRNA. [11][12][13][14][15][16] Antibody is an immune system-related biomolecule that can bind a speci c region on an antigen.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Ligation of Antibody and Cell-Penetrating Peptide for Efficient and Cell-Specific siRNA Delivery

Ando

Nakazawa

Miura

et al. 2021

Preprint

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A fusion protein comprising an antibody and a cell-penetrating peptide is a candidate molecule capable of efficient and cell-specific delivery of siRNA into cells in order to reduce the side effects of nucleic acid drugs. However, their expression in bacterial hosts, required for their development, often fails, impeding research progress. In this study, we separately prepared anti-EGFR nanobodies with the K-tag sequence MRHKGS at the C-terminus and arginine-9 (R9) with the Q-tag sequence LLQGS at the N-terminus, and enzymatically ligated them in vitro by microbial transglutaminase to generate Nanobody-R9, which is not expressed as a fused protein in E. coli. Nanobody-R9 exhibited a maximum reaction efficiency of 85.1%, without changing the properties of the nanobody or R9. Nanobody-R9 successfully delivered siRNA into the cells, and the cellular influx of siRNA increased with increase in the ratio of Nanobody-R9 to siRNA. We further demonstrated that the Nanobody-R9–siRNA complex, at a 30:1 ratio, induced RNAi of target mRNA with approximately 52% efficiency compared to lipofectamine.

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Section: Introductionmentioning

confidence: 99%

Enzymatic Ligation of Antibody and Cell-Penetrating Peptide for Efficient and Cell-Specific siRNA Delivery

Ando

Nakazawa

Miura

et al. 2021

Preprint

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“…Although RNAi therapy is promising, siRNAs delivered in vivo face significant barriers, including serum instability, renal excretion, rapid uptake by the reticuloendothelial system, lack of cell/tissue-type specificity, poor intracellular delivery, and endosomal entrapment [4–6]. Therefore, strategies using direct chemical modification [7–9] and viral [10–12] / non-viral [13,14] delivery systems have been explored to enable intracellular accumulation of siRNAs.…”

Section: Introductionmentioning

confidence: 99%

Dual peptide-mediated targeted delivery of bioactive siRNAs to oral cancer cells in vivo

et al. 2017

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Objectives Despite significant advances in cancer treatment, the prognosis for oral cancer remains poor in comparison to other cancer types, including breast, skin, and prostate. As a result, more effective therapeutic modalities are needed for the treatment of oral cancer. Consequently, in the present study, we examined the feasibility of using a dual peptide carrier approach, combining an epidermal growth factor receptor (EGFR)-targeting peptide with an endosome-disruptive peptide, to mediate targeted delivery of small interfering RNAs (siRNAs) into EGFR-overexpressing oral cancer cells and induce silencing of the targeted oncogene, cancerous inhibitor of protein phosphatase 2A (CIP2A). Materials and methods Fluorescence microscopy, real-time PCR, Western blot analysis, and in vivo bioimaging of mice containing orthotopic xenograft tumors were used to examine the ability of the dual peptide carrier to mediate specific delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells/tissues. Results Co-complexation of the EGFR-targeting peptide, GE11R9, with the endosome-disruptive 599 peptide facilitated the specific uptake of siRNAs into oral cancer cells overexpressing EGFR in vitro with optimal gene silencing observed at a 60:30:1 (GE11R9:599:siRNA) molar ratio. Furthermore, when administered systemically to mice bearing xenograft oral tumors, this dual peptide complex mediated increased targeted delivery of siRNAs into tumor tissues in comparison to the 599 peptide alone and significantly enhanced CIP2A silencing. Conclusion Herein we provide the first report demonstrating the clinical potential of a dual peptide strategy for siRNA-based therapeutics by synergistically mediating the effective targeting and delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells.

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