Intracellular Gene Transfer in Rats by Tail Vein Injection of Plasmid DNA

Abstract

Abstract. In this study, we examined the effect of various factors on gene delivery efficiency of tail vein injection of plasmid DNA into rats. We measured the level of reporter gene expression in the internal organs including the lung, heart, spleen, kidney, and liver as function of injection volume, injection time, and DNA dose. Persistency of reporter gene expression in transfected animals was also examined. We demonstrated that plasmid delivery to rats by the tail vein is effective as long as the volume of injected DNA solution is adjusted to 7-8% of body weight with an injection time of less than 10 s. With the exception of a short-term increase in serum concentration of alanine aminotransferase and transient irregularity in cardiac function during and soon after the injection, the procedure is well tolerated. Lac Z staining of the liver from transfected animals showed approximately 5-10% positive cells. Persistency test for transgene expression in animals using plasmid carrying cDNA of human alpha 1 antitrypsin gene driven by chicken beta actin gene promoter with CMV enhancers showed peak level of transgene product 1 day after the injection followed by a gradual decline with time. Peak level was regained by a second injection performed on day 38 after the first injection. These results show that tail vein injection is an effective means for introducing plasmid DNA into liver cells in rats. We believe that this procedure will be extremely useful for gene function studies in the context of whole animal in rats.

Hydrodynamic cell delivery for simultaneous establishment of tumor growth in mouse lung, liver and kidney

“…[4][5][6][7] The hydrodynamic delivery method is a method developed for delivery of either DNA or siRNA for gene function and gene therapy studies in rodents. [8][9][10][11] It involves a rapid tail-vein injection of a large volume (equal to 8-10% body weight) of siRNA/ DNA solution into a mouse or rat to build high intravascular pressure, consequently, increasing endothelia permeability and…”

A method to facilitate and monitor expression of exogenous genes in the rat kidney using plasmid and viral vectors

“…By impacting fluid pressures within thin and stretchable capillaries (37), this process is believed to increase the permeability of the capillary endothelium (37) and epithelial junctions (44), and to generate transient pores in plasma membranes facilitating the cellular internalization of macromolecules of interest (17). Systemic hydrodynamic gene delivery has been used successfully with the liver as the target organ (5,37,49), but renal expression has not been achieved with systemic delivery. However, the unique anatomy of the kidney provides various innate delivery paths (renal artery, renal vein, and ureter) that may be ideal for hydrodynamic gene delivery (47) directly to the organ.…”

“…In particular, it has been suggested that the relatively low-pressure renal venous system would provide a feasible site for hydrodynamic injections, by facilitating exogenous transgene delivery and limiting injection-related tissue damage (38,47). Plasmids (7,16,25,30,31,36,(45)(46)(47)49), baculovirus (1,2,23,24,39), and adenovirus (21,29,35,41,45) vectors have been previously identified as useful agents for in vitro and in vivo mammalian gene delivery. These vectors are reported to generate relatively low levels of pathogenicity and toxicity, are capable of carrying large DNA inserts, and can be generated in sufficiently high concentrations to provide sustained transgene expression (22,23,29,30).…”

Hydrodynamic Delivery

“…Experimentally, a 27-gauge needle is commonly used in tail vein injections in mice (Liu et al, 1999), whereas 20-24 gauge needles are appropriate for rats depending on body weight (Zhou, Kamimura, Zhang, & Liu, 2010). Other sized needles can also be used depending on the injection volume and speed and the type of blood vessel receiving the injection.…”

“…For maximal delivery efficiency, mice should be injected as rapidly as possible, typically in 3-5 s (Liu et al, 1999). Instead, rats are typically injected at a rate of 2 ml/s using a power injector , although a handheld syringe can be used (Zhou et al, 2010). The isolated liver of the rabbit was injected at a rate of 15-20 ml/s, which can only be accomplished using a power injector (Eastman et al, 2002).…”