Dataset Information

Effect of the combination of cationic lipid and phospholipid on gene-knockdown using siRNA lipoplexes in breast tumor cells and mouse lungs.

ABSTRACT: Previously, using three types of cationic lipids, the effect of phospholipids in liposomal formulations on gene-knockdown efficacy was determined after in vitro and in vivo transfection with small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) containing various cationic lipids and phospholipids. In the present study, six other types of cationic lipids, namely N,N-dimethyl-N-tetradecyltetradecan-1-aminium bromide, N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16), 2-[bis{2-(tetradecanoyloxy)ethyl}amino]-N,N,N-trimethyl-2-oxoethan-1-aminium chloride (DC-6-14), 1,2-di-O-octadecenyl-3-trimethylammonium propane chloride (DOTMA), 1,2-distearoyl-3-trimethylammonium-propane chloride (DSTAP) and 1,2-dioleoyl-3-dimethylammonium-propane were selected, and the effect of phospholipids in liposomal formulations containing each cationic lipid on gene-knockdown was evaluated. A total of 30 types of cationic liposomes composed of each cationic lipid with phosphatidylethanolamine containing unsaturated or saturated diacyl chains (C14, C16 or C18) were prepared. Regardless of the type of cationic lipid, the inclusion of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) in the liposomal formulations resulted in injectable size of siRNA lipoplexes after mixing of siRNA and cationic liposomes. Transfection of their lipoplexes with luciferase (Luc) siRNA into human breast cancer MCF-7-Luc cells stably expressing Luc led to a strong knockdown of Luc. Furthermore, the systemic injection of siRNA lipoplexes composed of DC-1-16, DC-6-14, DOTMA or DSTAP with DOPE resulted in siRNA accumulation in the lungs. Significant gene-knockdown was observed in the lungs of mice following the systemic injection of siRNA lipoplexes containing DC-1-16 and DOPE. Cationic liposomes composed of DC-1-16 and DOPE serve as potential carriers for in vitro and in vivo siRNA transfection.

SUBMITTER: Hattori Y

PROVIDER: S-EPMC10463231 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Effect of the combination of cationic lipid and phospholipid on gene-knockdown using siRNA lipoplexes in breast tumor cells and mouse lungs.

Hattori Yoshiyuki Y Tang Min M Aoki Aya A Ezaki Momoka M Sakai Hana H Ozaki Kei-Ichi KI

Molecular medicine reports 20230818 4

Previously, using three types of cationic lipids, the effect of phospholipids in liposomal formulations on gene-knockdown efficacy was determined after in vitro and in vivo transfection with small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) containing various cationic lipids and phospholipids. In the present study, six other types of cationic lipids, namely N,N-dimethyl-N-tetradecyltetradecan-1-aminium bromide, N-hexadecyl-N,N- ...[more]

PMID: 37594053

Similar Datasets

Project description:Previously, it was reported that mRNA/cationic liposome complexes (mRNA lipoplexes) composed of the cationic triacyl lipid, 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl)propan-2-yl)amino)-N,N,N- trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12), with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine and poly(ethylene glycol) cholesteryl ether, induce high protein expression in human cervical carcinoma HeLa cells. In the present study, the authors aimed to optimize mRNA transfection using TC-1-12-based mRNA lipoplexes. mRNA lipoplexes were prepared at various charge ratios (+:-) using modified ethanol injection (MEI) and thin-film hydration (TFH) methods and compared the protein expression efficiency after transfection of HeLa cells with the developed mRNA lipoplexes. Firefly luciferase (FLuc) and enhanced green fluorescent protein (EGFP) mRNA lipoplexes prepared using the MEI method exhibited higher Luc and EGFP expression levels in cells than those prepared using the TFH method. Moreover, FLuc mRNA lipoplexes prepared using the MEI and TFH methods at charge ratios of 3:1 and 4:1, respectively, exhibited the highest Luc expression in cells. However, transfection with mRNA lipoplexes using the MEI and TFH methods induced moderate cytotoxicity in HeLa cells (46 and 57% cell viability, respectively). Furthermore, Cy5-labeled mRNA lipoplexes, which were prepared using the MEI method, showed higher cellular uptake of mRNA than those prepared using the TFH method. In the transfection of FLuc mRNA lipoplexes prepared using the MEI method, the storage of the lipid-ethanol solution at 37˚C for 4 months did not decrease Luc expression in HeLa cells. Additionally, FLuc mRNA lipoplexes prepared using the MEI method, induced relatively high Luc expression in human prostate carcinoma PC-3 and human liver cancer HepG2 cells with low cytotoxicity (103 and 81% cell viability, respectively). Overall, the results highlighted the potential of TC-1-12-based mRNA lipoplexes prepared using the MEI method for efficient mRNA delivery to cells.

Dataset Information

Effect of the combination of cationic lipid and phospholipid on gene-knockdown using siRNA lipoplexes in breast tumor cells and mouse lungs.

Publications

Effect of the combination of cationic lipid and phospholipid on gene-knockdown using siRNA lipoplexes in breast tumor cells and mouse lungs.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets