Transient expression of proteins by hydrodynamic gene delivery in mice.
Efficient expression of transgenes in vivo is of critical importance in studying gene function and developing treatments for diseases. Over the past years, hydrodynamic gene delivery (HGD) has emerged as a simple, fast, safe and effective method for delivering transgenes into rodents. This technique relies on the force generated by the rapid injection of a large volume of physiological solution to increase the permeability of cell membranes of perfused organs and thus deliver DNA into cells. One of the main advantages of HGD is the ability to introduce transgenes into mammalian cells using naked plasmid DNA (pDNA). Introducing an exogenous gene using a plasmid is minimally laborious, highly efficient and, contrary to viral carriers, remarkably safe. HGD was initially used to deliver genes into mice, it is now used to deliver a wide range of substances, including oligonucleotides, artificial chromosomes, RNA, proteins and small molecules into mice, rats and, to a limited degree, other animals. This protocol describes HGD in mice and focuses on three key aspects of the method that are critical to performing the procedure successfully: correct insertion of the needle into the vein, the volume of injection and the speed of delivery. Examples are given to show the application of this method to the transient expression of two genes that encode secreted, primate-specific proteins, apolipoprotein L-I (APOL-I) and haptoglobin-related protein (HPR).
Department of Biological Sciences, Hunter College, CUNY.
Daniella
D
Kovacsics
Department of Biological Sciences, Hunter College, CUNY; raper@genectr.hunter.cuny.edu.
Jayne
J
Raper
D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D005260:Female; D057446:Hydrodynamics; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D019076:Transgenes
D011506:Proteins; D004247:DNA