1. Transfection can be accomplished through the use of cationic polymers such as DEAE-dextran or polyethylenimine. The negatively charged DNA binds to the polycation and the complex is taken up by the cell via endocytosis. After endocytosis, the DNA is released by the endosome into the cytosol. Many cells can be transfected much more efficiently with polymers! www.cmu.edu Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Polymer-based In Vivo Transfection Kit

2. Some polymers have been developed that are capable of forming complexes with DNA or RNA and have the potential to form micelles. The transfection reaction is usually performed under aqueous conditions which enables the lipophilic portion of the amphiphilic compound to form the micelle core within which the exogenous nucleic acids are ensconced. Superior transfection efficiencies can be achieved in cell lines that are refractory to liposome-based transfection. All Altogen® In Vivo Transfection Kits supplied with ready-to-run transfection protocols that eliminate the need for extensive transfection optimization experiments Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Products > Polymer-based In Vivo Transfection Kit About

3. www.mdpi.org Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Products > Polymer-based In Vivo Transfection Kit Mechanism of Action

4. Biodegradable polymer-based in vivo reagent No detectable inflammatory response Efficient delivery to the lung, liver, kidney, and spleen via systemic administration Efficient siRNA and plasmid DNA delivery via direct subcutaneous tumor injection (multiple tumor types) Functionally tested in mice Applicable for plasmid DNA/siRNA co-injection Modes of administration: Systemic intravenous (i.v.) injection Direct intratumoral (i.t.) injection Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Products > Polymer-based In Vivo Transfection Kit Product Details

5. Figure 2. Intratumoral administration (i.t.) of Polymer-based In Vivo reagent conjugated with siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues were collected and RNA isolated 48 hours after first injection. Samples were analyzed by qRT-PCR for Lamin A/C gene expression levels. Ribosomal RNA levels were used to normalize the Lamin A/C data. Data are means ± SD (n=6). Products > Polymer-based In Vivo Transfection Kit Data

6. Figure 2. Intratumoral administration (i.t.) of Polymer-based In Vivo reagent conjugated with siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues were collected and RNA isolated 48 hours after first injection. Samples were analyzed by qRT-PCR for Lamin A/C gene expression levels. Ribosomal RNA levels were used to normalize the Lamin A/C data. Data are means ± SD (n=6). Products > Polymer-based In Vivo Transfection Kit Data

7. Products > Polymer-based In Vivo Transfection Kit Data Figure 3. Systemic administration (i.v.) of Polymer In Vivo Transfection Reagent conjugated with 80 ug of chemically modified siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues (Liver, Brain, Muscle, Kidney, Tumor, Heart, Spleen, Lung) were collected and total protein fraction isolated 72 hours after first injection. Samples were analyzed by Western Blot Analysis for Lamin A/C gene expression levels.

8. Products > Polymer-based In Vivo Transfection Kit Data Figure 4. Systemic administration (i.v.) of Polymer In Vivo Transfection Reagent conjugated with 80 ug of chemically modified siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues (Liver, Brain, Muscle, Kidney, Tumor, Heart, Spleen, Lung) were collected and total protein fraction isolated 72 hours after first injection. Samples were analyzed by Western Blot Analysis for Lamin A/C gene expression levels.

9. Products > Polymer-based In Vivo Transfection Kit Data Figure 5. Systemic administration (i.v.) of Polymer In Vivo Transfection Reagent conjugated with 80 ug of chemically modified siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues (Liver, Brain, Muscle, Kidney, Tumor, Heart, Spleen, Lung) were collected and total protein fraction isolated 72 hours after first injection. Samples were analyzed by Western Blot Analysis for Lamin A/C gene expression levels.

10. Products > Polymer-based In Vivo Transfection Kit Data Figure 6. Systemic administration (i.v.) of Polymer In Vivo Transfection Reagent conjugated with 80 ug of chemically modified siRNA targeting Lamin A/C mRNA or non-silencing control siRNA following the recommended protocol. Tissues (Liver, Brain, Muscle, Kidney, Tumor, Heart, Spleen, Lung) were collected and total protein fraction isolated 72 hours after first injection. Samples were analyzed by Western Blot Analysis for Lamin A/C gene expression levels.

11. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Contents and Shipping: POLYMER-based in vivo transfection reagent is supplied in liquid form at a concentration of 3 mg/ml. Transfection Enhancer is supplied in liquid form at a concentration of 1 mg/ml. Shipped at ambient temperature. Description: POLYMER transfection reagent is a biodegradable polymer-based formulation optimized for in vivo delivery of small RNA (siRNA, shRNA, microRNA) and plasmid DNA. POLYMER in vivo transfection reagent is a proprietary, animal-origin-free formulation. Storage: Store reagent at 4ºC upon receipt. If stored properly, reagent is stable for 6 months. Product Qualification: Reagents are tested for functional activity, absence of nuclease contamination and microbial contamination. Reagent Properties: Polymer conjugated complexes are stable in serum for at least 16 hours. Efficient delivery to the liver, lung, kidney, and certain tumor types via systemic administration. Efficient siRNA and plasmid DNA delivery via direct subcutaneous tumor injection (various tumor types). Minimal toxicity. Products > Polymer-based In Vivo Transfection Kit General Protocol

12. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Biodegradable polymer-based in vivo reagent No detectable inflammatory response Efficient delivery to the lung, liver, kidney, and spleen via systemic administration Efficient siRNA and plasmid DNA delivery via direct subcutaneous tumor injection (multiple tumor types) Functionally tested in mice Applicable for plasmid DNA/siRNA co-injection Modes of administration: Systemic intravenous (i.v.) injection Direct intratumoral (i.t.) injection Products > Polymer-based In Vivo Transfection Kit Summary