Open AccessCalcium based siRNA coating: a novel approach for knockdown of HER2 gene in MCF-7 cells using gold nanoparticles
Author(s) -
Abdul Sami,
S.M. Saqalan Naqvi,
Mazhar Qayyum,
Abida Raza Rao,
Uteuliyev Yerzhan Sabitaliyevich,
Muhammad Sheeraz Ahmad
Publication year - 2020
Publication title -
cellular and molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.371
H-Index - 71
eISSN - 1165-158X
pISSN - 0145-5680
DOI - 10.14715/cmb/2020.66.6.19
Subject(s) - transfection , gene knockdown , microbiology and biotechnology , cytotoxicity , surface modification , chemistry , small interfering rna , rna interference , gene delivery , housekeeping gene , colloidal gold , mcf 7 , calcium , gene expression , biophysics , in vitro , cancer cell , gene , materials science , nanoparticle , rna , biochemistry , biology , nanotechnology , cancer , human breast , genetics , organic chemistry
Surface functionalization of nanoparticles (NPs) for therapeutic siRNA delivery into cancer cells has gained interest. The present study was designed for surface functionalization of gold nanoparticles (AuNPs) for efficient siRNA delivery and knockdown in cancer cells. In order to achieve this objective, AuNPs were coated with HER2-siRNA in the presence of 11-mercaptoundecanoic acid (11-MUA), calcium chloride (CaCl2) and polyethyleneimine (PEI) in alternate charge bearing successive layers. MCF-7 cells were cultured and transfected with fabricated assembly of AuNPs. Cytotoxicity analysis revealed that the half inhibitory concentration (IC50) for the formulation was 45.35 nM . Total RNA was isolated from transfected cells, reverse transcribed into complementary DNA (cDNA) and real-time polymerase chain reaction (RT-PCR) was performed. The RT-PCR based delta-delta Ct analysis in treated cells revealed a significant 18.94 times decrease (p<0.001) in the expression of HER2 gene standardized with ACTB housekeeping gene as compared to untreated cells, which makes this formulation a potent approach for siRNA delivery and gene knockout.