DOTMA:DOPE liposomes consist of DOTMA, a cationic lipid, and DOPE, a neutral helper lipid. This composition is specifically designed to enhance the delivery of nucleic acids, such as DNA and RNA, into cells by neutralizing the negative charge of the nucleic acids through electrostatic interactions, thereby facilitating their cellular uptake.
Nucleic acids can be efficiently loaded into DOTMA:DOPE liposomes through a process known as complexation, where the negatively charged nucleic acids interact electrostatically with the positively charged liposomes. This process is usually performed at room temperature, with careful titration of nucleic acid to achieve the desired charge ratio.
The pH sensitivity of DOTMA:DOPE liposomes is crucial for their gene delivery capabilities. Upon encountering the acidic environment inside endosomes, these liposomes undergo a pH-dependent conformational change that promotes endosomal membrane destabilization and subsequent release of their genetic cargo into the cytoplasm, which is a critical step for successful gene expression.
Membrane fusion is a key mechanism through which DOTMA:DOPE liposomes deliver their cargo into cells. After endocytosis, the fusogenic property of DOPE facilitates the fusion of liposomes with endosomal membranes, leading to the release of the encapsulated nucleic acids into the cytoplasm, where they can exert their biological effects.
The transfection efficiency of DOTMA:DOPE liposomes can be evaluated using a reporter gene assay, such as luciferase or GFP, where the expression level of the reporter gene in the target cells indicates the success of nucleic acid delivery. Quantitative PCR and flow cytometry are also commonly used methods to assess transfection efficiency.
Confocal images of primary cells transfected with epithelial targeting peptide-modified liposomes (DOTMA:DOPE), α-ENaC (green), actin (red) and DAPI (blue) to label the nuclei.
This study investigated the efficacy of epithelial targeting peptide-modified liposomes (DOTMA:DOPE) for siRNA delivery to treat cystic fibrosis (CF). ENaC hyperactivity and dysregulation are associated with respiratory dysfunction, pulmonary pathology, and dehydration of respiratory secretions in CF. Researchers evaluated the silencing efficiency of targeting peptide-modified DOTMA:DOPE liposomes as carriers for α-ENaC siRNA. Confocal microscopy demonstrated a significant decrease in fluorescence staining of α-ENaC. This indicates that targeting peptide-modified DOTMA:DOPE liposomes are a reliable and safe non-viral siRNA delivery system for the treatment of CF.
Manunta, Maria DI, et al. "Delivery of ENaC siRNA to epithelial cells mediated by a targeted nanocomplex: a therapeutic strategy for cystic fibrosis." Scientific reports. 7.1 (2017): 700. Under Open Access license CC BY 4.0, the image was edited from figure 5.
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