Solid-phase synthesis, characterization and RNAi activity of branch and hyperbranch siRNAs
- Resource Type
- Authors
- Brittany A. Blackman; Christopher J. Parronchi; Allan D. Blake; Anthony Maina; David Sabatino; Maria E. Bender; Eva L. Morozko
- Source
- Bioorganic & Medicinal Chemistry Letters. 23:5270-5274
- Subject
- Thermal denaturation
Small interfering RNA
Programmed cell death
Circular dichroism
Cell Survival
Clinical Biochemistry
Pharmaceutical Science
Antineoplastic Agents
Biochemistry
Solid-phase synthesis
RNA interference
Drug Discovery
Humans
RNA, Small Interfering
Endoplasmic Reticulum Chaperone BiP
Molecular Biology
Solid-Phase Synthesis Techniques
Microscopy, Confocal
Chemistry
Organic Chemistry
Hep G2 Cells
Molecular biology
Cancer cell
Molecular Medicine
Cancer gene
RNA Interference
- Language
- ISSN
- 0960-894X
Linear, branch and hyperbranch siRNAs were effectively prepared for down-regulating GRP78 expression and inducing cell death in HepG2 liver cancer cells. Branch and hyperbranch GRP78 siRNAs were synthesized by automated solid-phase synthesis in good yields (44–78%) and isolated in excellent purities (>99%) following HPLC purification. Moreover, siRNAs adopted stable intramolecular hybrids as discerned by native PAGE and thermal denaturation studies. These sequences also exhibited the pre-requisite A-type helical trajectory for triggering RNAi activity as determined by CD spectroscopy. Biological studies confirmed potent suppression of GRP78 expression (50–60%) while compromising cancer cell viability by ∼20%. Thus, branch and hyperbranch siRNAs may serve as potent siRNA candidates in cancer gene therapy applications.