DDAB Liposomes for DNA/RNA Delivery

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DDAB Liposomes form stable complexes with DNA, RNA, oligonucleotides, and proteins. BOC Sciences offers a number of popular cationic liposome reagents made from popular cationic lipids prepared under sterile/low endotoxin conditions with nuclease-free water for the highest quality. The LUVs obtained have a small particle size and are best suited for forming small and stable lipid complexes with nucleic acids.

What are DDAB Liposomes?

DDAB (dimethyldioctadecylammonium bromide) liposomes are cationic liposomes for DNA/RNA delivery. Liposomes have a spherical lipid bilayer structure that encapsulates and protects nucleic acids (such as DNA and RNA) from degradation and improves their delivery to the cell. DDAB Liposomes consist of positively charged lipids that interact with negatively charged nucleic acids to facilitate their encapsulation in the liposome. Studies have shown that DDAB liposomes can efficiently deliver DNA/RNA to a wide range of cell types, including cells that are difficult to transfect. They are also used for various applications such as gene therapy, drug delivery and vaccine development.

Features of DDAB Liposomes for DNA/RNA Delivery

Non-cholesterol cationic lipids for gene delivery
pH-sensitive liposomes
No accumulation or degradation

DNA/RNA Delivery Mechanism

DDAB liposomes can deliver nucleic acids into cells either by endocytosis or through cell membrane fusion. Fused cationic liposomes deliver nucleic acids almost exclusively through cell membrane fusion. Fusion between positively charged cationic liposomes and negatively charged cell surfaces effectively delivers DNA directly across the plasma membrane. This process bypasses the endosomal-lysosomal pathway, leading to degradation of anionic liposome preparations. Cationic liposomes in the laminar phase deliver nucleic acids via endocytosis, specifically lattice protein-mediated endocytosis, cisternae-mediated endocytosis (CavME), and megacytosis.

How are DDAB Liposomes Prepared?

DDAB liposomes are manufactured similarly to liposomes. There are a variety of methods that can be used to form cationic liposomes, such as sonication, extrusion, and vortexing. However, the shear forces associated with these methods are capable of destroying nucleic acids prior to encapsulation. Microfluidics is a more suitable preparation that aims to form cationic liposomes without the shear forces and damage associated with existing methods.

How to Assess the Effectiveness of DDAB Liposomal Gene Delivery?

(1) Target Gene Expression

When DDAB liposomes deliver DNA/RNA encoding an expression reporter or a specific protein, their effectiveness is assessed.

(2) Fluorescent Dye Labeling

When DNA/RNA or vectors are conjugated with fluorescent dye markers, the fluorescent signal inside the cell can be observed by fluorescence microscopy. DDAB liposomes have delivered DNA/RNA into the cell if the fluorescent signal is visible and localized in the nucleus or cytoplasm.

(3) PCR/RT-PCR Analysis

Target genes were detected by PCR/RT-PCR, which is a polymerase chain reaction-based method. In order to assess the effect of DDAB liposome delivery, cellular RNA/DNA was extracted and then amplified using specific primers. This was then compared with the number and size of amplification products obtained before and after DDAB liposome delivery.

(4) Flow Cytometry

The effect of delivery can be quantitatively analyzed using cell surface markers or internal markers. The efficiency of DDAB liposome delivery is assessed by measuring the fluorescent signal or the expression level of specific markers in the cells.