TransIT®-QR Delivery Solution and Starter Kit

Figure 1. TransIT-QR Delivery Solution Effectively Delivers the Label IT® Cy™3 RNAi Delivery Control to Hepatocytes. TransIT-QR 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) 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 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 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.