Solid-phase synthesis, characterization and RNAi activity of branch and hyperbranch siRNAs

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...

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Bibliographic Details
Published in:Bioorganic & medicinal chemistry letters 2013-10, Vol.23 (19), p.5270-5274
Main Authors: Maina, Anthony, Blackman, Brittany A., Parronchi, Christopher J., Morozko, Eva, Bender, Maria E., Blake, Allan D., Sabatino, David
Format: Article
Language:English
Subjects:
Antineoplastic Agents - chemical synthesis
Antineoplastic Agents - chemistry
Antineoplastic Agents - pharmacology
Branch siRNA
Cell death
Cell Survival - drug effects
cell viability
circular dichroism spectroscopy
denaturation
gene therapy
GRP78 silencing
Hep G2 Cells
high performance liquid chromatography
Humans
Hyperbranch siRNA
liver neoplasms
Microscopy, Confocal
Oncogene therapy
polyacrylamide gel electrophoresis
RNA Interference
RNA, Small Interfering - chemical synthesis
RNA, Small Interfering - genetics
RNA, Small Interfering - pharmacology
RNAi
small interfering RNA
Solid-Phase Synthesis Techniques
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Summary: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.
ISSN:0960-894X
1464-3405
DOI:10.1016/j.bmcl.2013.08.033