1. Nanoparticles are particles between 1 and 100 nanometers in size. In nanotechnology, a particle is defined as a small object that behaves as a whole unit with respect to its transport and properties. Through the Altogen® Biosystems Nanoparticle-based Transfection Reagent Kit, it is possible to ensure efficient delivery to the heart, lung, liver, pancreas, kidney, and multiple tumor types via systemic administration and functionally tested in mice. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products www.wikipedia.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 > Nanoparticle-based In Vivo Transfection Kit

2. All Altogen® In Vivo Transfection Kits supplied with ready-to-run transfection protocols that eliminate the need for extensive transfection optimization experiments Nanoparticle-based in vivo transfection reagent is a proprietary, animal- origin-free formulation of chemically engineered nanoparticles optimized for siRNA, shRNA, miRNA, and plasmid DNA in vivo delivery. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Products > Nanoparticle-based In Vivo Transfection Kit About

3. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Nanoparticles can be used as carriers for nucleic acids. The nanoparticles have an approximate size of 100 nm and are additionally coated with biological polymers to allow loading of nucleic acids. Particles and nucleic acids form complexes by ionic interaction of the negatively charged nucleic acid and the positively charged surface of the nanoparticle. The binding of the negatively charged nucleic acids (RNA, DNA, mRNA, siRNA) to the positively charged iron particles occurs relatively fast, and within 30 minutes. After complex formation, the loaded particles are administered in vivo. The nanoparticles are accumulated in target tissues. Over time, the nanoparticles are degraded and the iron enters the normal iron metabolism. Products > Nanoparticle-based In Vivo Transfection Kit Mechanism of Action

4. Nanoparticle-based reagent Complexes are stable in serum for 16 hours Efficient delivery to the heart, lung, liver, pancreas, kidney, and multiple tumor types via systemic administration Efficient siRNA and plasmid DNA delivery via IV, IT, or direct subcutaneous tumor injection (various tumor types) Minimal toxicity Functionally tested in mice and rats Applicable for plasmid DNA/siRNA co-injection Developed and manufactured by Altogen Biosystems 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 > Nanoparticle-based In Vivo Transfection Kit Product Details

5. Non-lipid formula that allows for stable complexes for injection. Entirely free of serum and protein of animal origin. Our kits are easy to use and effective: Mix, incubate, dilute, and inject! Our kits allow for sustained knockdown after a single injection! Target various organs through different administration routes. There is very low in vivo toxicity and it is safe to use as there is no detectable inflammatory response. Used for preclinical research worldwide! Products > Nanoparticle-based In Vivo Transfection Kit Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

6. The Altogen® Biosystems Nanoparticle-based In Vivo Transfection Kit is developed and manufactured exclusively by Altogen® Biosystems and allows for very efficient In Vivo delivery via tissue-targeting, outlined further in the next slide and in our easy-to-use protocols that optimize efficiency. Products > Nanoparticle-based In Vivo Transfection Kit Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

7. Combine diluted exogenous nucleic acid material and diluted Altogen® Biosystems Nanoparticle-based Transfection Reagent, vortex, incubate and choose delivery system for targeted tissue. The experimental animal recipient, the targeted tissue and route of administration are the factors that will determine the amount of exogenous nucleic acids and suggested injection volume. Products > Nanoparticle-based In Vivo Transfection Kit Tissue-Targeting Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

8. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Altogen® Biosystems Nanoparticle-based Transfection Kit compares very favorably to other commercially available kits Tested for delivery in both rats and mice The hands-on time is less than 15 minutes! Successfully delivers maximum amount of siRNA, miRNA, DNA in vivo Altogen® Biosystems has proven results with a simple, quick and effective protocol. Products > Nanoparticle-based In Vivo Transfection Kit Product Details

9. Figure 1. Systemic administration (i.v.) of Liposome-PEG 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 > Nanoparticle-based In Vivo Transfection Kit Data

10. Figure 2. Intratumoral administration (i.t.) of Nanoparticle-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 > Nanoparticle-based In Vivo Transfection Kit Data

11. Products > Nanoparticle-based In Vivo Transfection Kit Data Figure 3. Systemic administration (i.v.) of Nanoparticle 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.

12. Products > Nanoparticle-based In Vivo Transfection Kit Data Figure 4. Systemic administration (i.v.) of Nanoparticle 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.

13. Products > Nanoparticle-based In Vivo Transfection Kit Data Figure 5. Systemic administration (i.v.) of Nanoparticle 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.

14. Products > Nanoparticle-based In Vivo Transfection Kit Data Figure 6. Systemic administration (i.v.) of Nanoparticle 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.

15. 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: Nanoparticle-based in vivo transfection reagent is supplied in liquid form at a concentration of 2.4 mg/ml. Transfection Enhancer is supplied in liquid form at a concentration of 1 mg/ml. Shipped at ambient temperature. 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: Nanoparticle conjugated complexes are stable in serum for at least 16 hours. Efficient delivery to the lung, liver, pancreas, kidney, and certain tumor types via systemic administration. Efficient siRNA and plasmid DNA delivery via direct subcutaneous tumor injection (various tumor types). Minimal toxicity. General Guidelines: Handle animals and conduct experiments according to national regulations and approvals by the local IACUC and ethics committee. Please note that mouse tail vein injection may require skills, experience, and associated equipment (e.g. restraining device). All persons handling animals should be properly trained. Products > Nanoparticle-based In Vivo Transfection Kit General Protocol

16. Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com Nanoparticle-based reagent Complexes are stable in serum for 16 hours Efficient delivery to the heart, lung, liver, pancreas, kidney, and multiple tumor types via systemic administration Efficient siRNA and plasmid DNA delivery via direct subcutaneous tumor injection (various tumor types) Minimal toxicity Functionally tested in mice Applicable for plasmid DNA/siRNA co-injection Developed and manufactured by Altogen Biosystems Products > Nanoparticle-based In Vivo Transfection Kit Summary