Publication
Development of lipofection vectors based in novel serine-based cationic gemini surfactants finely tuned by monoolein for therapeutic siRNA delivery
| Summary: | Gene therapy based on gene silencing with siRNA has evolved and gained great importance over the last few years as a method with great potential in the treatment of genetic diseases. However, a major challenge to the use of RNAi-based therapies is their efficient delivery through an appropriate vector. Cationic liposomes are an example of the most versatile nonviral vectors for the delivery of nucleic acids and have been researched in the past few years as a strategy to promote effective transfection of genetic material. This project has focused on the development and characterization of novel vectors for therapeutic siRNA delivery based in three different gemini amino acid-based surfactant derived from serine with an helper lipid called monoolein (MO). Monoolein lipid is a known promoter of inverted nonbilayer structures such as inverted cubic mesophases and could enhance the fusogenicity of serine-based gemini aggregates while modulating the properties of the final aggregates. The use of these vectors aims at therapeutic purposes, and thus requires a full characterization of the formed systems. The vectors must have robust properties and the ability to ensure efficient and reliable performance. Therein, it is shown that the chemical nature of the spacer linker (amine, amide and ester) in serine-based gemini surfactant influences the properties of the aggregates formed and their complexes with nucleic acids. On the other hand, the inclusion of MO in liposomal or micellar formulations, as helper lipid, induces modifications on the morphology of the aggregates. Typically, this leads to a reduction in the mean diameter of particles, without the need of mechanic processes such as sonication or extrusion to increase their stability over time. The three gemini:MO systems tested have good ability to complex efficiently siRNA and they did not show significant levels of cytotoxicity. This compactation of siRNA leads to liposomal/micellar-siRNA complexes with optimal mean diameters, which could be used in vitro and in vivo. Lastly, the capacity to deliver siRNA and thus silence the target gene was also evaluated. The results obtained showed high percentages of down regulations, indicating that the gemini:MO formulations studied in this work are good candidates as lipofection vectors for RNAi-therapies. |
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| Main Authors: | Costa, Catarina Dias da |
| Subject: | Ciências Naturais |
| Year: | 2015 |
| Country: | Portugal |
| Document type: | master thesis |
| Access type: | restricted access |
| Associated institution: | Universidade do Minho |
| Language: | English |
| Origin: | RepositóriUM - Universidade do Minho |
| Summary: | Gene therapy based on gene silencing with siRNA has evolved and gained great importance over the last few years as a method with great potential in the treatment of genetic diseases. However, a major challenge to the use of RNAi-based therapies is their efficient delivery through an appropriate vector. Cationic liposomes are an example of the most versatile nonviral vectors for the delivery of nucleic acids and have been researched in the past few years as a strategy to promote effective transfection of genetic material. This project has focused on the development and characterization of novel vectors for therapeutic siRNA delivery based in three different gemini amino acid-based surfactant derived from serine with an helper lipid called monoolein (MO). Monoolein lipid is a known promoter of inverted nonbilayer structures such as inverted cubic mesophases and could enhance the fusogenicity of serine-based gemini aggregates while modulating the properties of the final aggregates. The use of these vectors aims at therapeutic purposes, and thus requires a full characterization of the formed systems. The vectors must have robust properties and the ability to ensure efficient and reliable performance. Therein, it is shown that the chemical nature of the spacer linker (amine, amide and ester) in serine-based gemini surfactant influences the properties of the aggregates formed and their complexes with nucleic acids. On the other hand, the inclusion of MO in liposomal or micellar formulations, as helper lipid, induces modifications on the morphology of the aggregates. Typically, this leads to a reduction in the mean diameter of particles, without the need of mechanic processes such as sonication or extrusion to increase their stability over time. The three gemini:MO systems tested have good ability to complex efficiently siRNA and they did not show significant levels of cytotoxicity. This compactation of siRNA leads to liposomal/micellar-siRNA complexes with optimal mean diameters, which could be used in vitro and in vivo. Lastly, the capacity to deliver siRNA and thus silence the target gene was also evaluated. The results obtained showed high percentages of down regulations, indicating that the gemini:MO formulations studied in this work are good candidates as lipofection vectors for RNAi-therapies. |
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