lipid mediated lipofection
TRANSCRIPT
Muhammad Bilal YusufStudent of Bs-Biotechnology at International
Islamic University Islamabad, Pakistan.Email: [email protected]
Lipid Mediated Lipofection
Objectives
Lipid Mediated Lipofection ????
Principle
Advantages
Disadvantages
Lipid Mediated LipofectionLipofection (or liposome transfection) is
a technique used to inject genetic material into a cell by means of liposomes.
which are vesicles that can easily merge with the cell membrane since they are both made of a phospholipid bilayer.
Lipofection generally uses a positively charged (cationic) lipid to form a structure with the negatively charged (anionic) genetic material.
Fusion of the liposome/nucleic acid transfection complex with the negatively charged cell membrane takes place.
The transfection complex is thought to enter the cell through endocytosis.
Once inside the cell, the complex must escape the endosomal pathway, diffuse through the cytoplasm, and enter the nucleus for gene expression.
Principle
AdvantagesYields high transfection efficiencies.Works in a wide variety of eukaryotic cells.Is simple to perform.Ensures consistently reproducible results.Moreover, a number of cell lines normally
resistant to transfection by other methods transfect successfully with cationic lipid reagents.
DisadvantagesQuite low transfection efficiency in
suspension cells. Dependence on cell division.As well as on high rate of endocytosis.Not Applicable to all cell types.
Problems with traditional methods
Methods like calcium phosphate co-precipitation, DEAE-dextran, polybrene, and electroporation include problems such as:
low efficiency of DNA deliverypoor reproducibilitycell toxicityinconvenience
