TransIT®-QR Delivery Solution and Starter Kit

A nontoxic, "Quick Recovery" in vivo delivery solution that delivers DNA or siRNA to the livers of laboratory mice using hydrodynamic tail vein injection

• Naked Nucleic Acid Delivery—Achieve higher levels of gene expression than compared to polylysine and PEE methods.
• Versatile Platform—for DNA or siRNA delivery using the hydrodynamic tail vein injection to the mouse strain of your choice.
• Low Toxicity—Minimized loss of cardiac output compared to saline injections, which allows for quick recovery (within minutes) of the mouse post-injection.

Kit Components:

Each TransIT-QR Starter Kit contains the TransIT-QR Hydrodynamic Delivery Solution, 10 syringes, needles and alcohol swabs, and a mouse restraint device designed specifically for tail vein injections. The TransIT-QR Hydrodynamic Delivery Solution is also sold separately as indicated above.

Data

Figure 1: TransIT-QR Solution Effectively Delivers the Label IT® Cy™3 RNAi Delivery Control to Hepatocytes.
Figure 2: Achieve Low Toxicity Using the TransIT-QR Hydrodynamic Delivery Solution.
Figure 3: Endogenous Target Knockdowns Are Achievable in the Liver Using Hydrodynamic Delivery of siRNA.
Figure 4: Knockdown of Multiple Organs After Co-delivery of siRNA and Target Plasmids.

Figure 1. TransIT-QR Hydrodynamic Delivery Solution Effectively Delivers the Label IT® Cy™3 RNAi Delivery Control to Hepatocytes. TransIT-QR Hydrodynamic Delivery Solution was used to deliver 25 µg of Label IT Cy™3 RNAi Delivery Control (red) to a mouse using hydrodynamic delivery via the tail vein. Forty-five minutes post-injection the liver was harvested and sections were fixed then counterstained to stain the nuclei (blue) and to stain the actin (green).

Figure 2. The TransIT-QR Solution Minimizes the Loss of Cardiac Output Compared to Injections Performed Using Normal Saline. Mice were anesthetized, connected to an arterial blood pressure monitor, and injected with either TransIT-QR (Quick Recovery) Hydrodynamic Delivery Solution (blue) or normal saline (gray) using the hydrodynamic tail vein injection procedure. The relative cardiac output was estimated by multiplying the heart rate by the pulse pressure (difference between systolic and diastolic pressures) and scaled to the pre-injection cardiac output.

Figure 3. Endogenous Target Knockdowns Are Achievable in the Liver Using Hydrodynamic Delivery of siRNA. Two different siRNAs directed against PPAR-alpha or a non-targeting control siRNA (40 µg each siRNA per mouse) were delivered via the hydrodynamic tail vein injection procedure to mice using the TransIT-QR Hydrodynamic Delivery Solution. Twenty-four hours post-injection, the livers were harvested and the PPAR-alpha levels were determined using qRT-PCR and averaged for each group.

Figure 4. Reporter Gene Knockdown in Multiple Organs After Co-delivery of siRNA and Reporter Plasmids. Mice were injected with 10 µg of plasmid encoding firefly luciferase, 1 µg of plasmid encoding Renilla luciferase, 5 µg of siRNA targeted against firefly luciferase, and 5 µg of control siRNA using the TransIT-QR Hydrodynamic Delivery Solution. Twenty-four hours later, the samples were assayed for firefly and Renilla luciferase activity. The level of firefly luciferase expression was normalized to Renilla activity and compared to numbers obtained using the control siRNA. Lewis et al., Nature Genetics 32: 107-8, 2002. NOTE: Following gene delivery via a tail vein injection, the highest level of transgene expression is found in the liver, with reduced levels of expression found in the spleen, lung, heart and kidneys. siRNA delivery to these organs is not sufficient to achieve appreciable endogenous targeted gene knockdown.

NOTE: In vivo hydrodynamic delivery of non-viral nucleic acids is covered by worldwide patents and patent applications of Mirus Bio Corporation, including U.S. Patent 6,627,616, U.S. Patent 6,379,966, and related filings worldwide. Purchase of this product does not provide a license to this delivery technology, which is required for all research and commercial uses by for-profit entities. To inquire about a license, please contact Mirus Bio Corporation.