Type of Submission
Poster
Keywords
Transfection methods, Calcium phosphate, lipofectamine, PCDH19, GFP
Proposal
Protocadherin-19 (PCDH19) is a protein coding gene expressed within the brains of vertebrate organisms. Mutations in this gene have been identified in human patients with PCDH19 clustering epilepsy, a form of epilepsy that primarily affects females and often coincides with varying degrees of intellectual disabilities. In addition, PCDH19 has been shown to play an important role in organizing cellular interactions in early neurodevelopmental processes. Ultimately, by gaining a greater understanding of the functions for PCDH19, we can further understand neurodevelopment and various neurodevelopmental diseases. For many in vitro studies of PCDH19, we first must obtain protein for use in the experiments. Human cell culture can be used to produces proteins of interest, when transfected with vectors containing the gene of interest. Here we tested two common transfection methods, lipofectamine transfection and calcium phosphate transfection, to determine which is the most cost effective method for the production of a PCDH19-GFP fusion protein. The cost per transfection is approximately five times higher per transfection with lipofectamine compared to calcium phosphate. However, the protein levels detected from lipofectamine were also consistently higher. The degree of increased protein production was variable and would require further replications to ascertain. From this data, we concluded that calcium phosphate is more cost effective, but lipofectamine may be necessary in some cases to reach sufficient levels of protein expression for some experiments.
Creative Commons License
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Cost Comparison of Calcium Phosphate Transfection and Lipofectamine Transfection for Production of PCDH19 Protein in HeLa Cells
Protocadherin-19 (PCDH19) is a protein coding gene expressed within the brains of vertebrate organisms. Mutations in this gene have been identified in human patients with PCDH19 clustering epilepsy, a form of epilepsy that primarily affects females and often coincides with varying degrees of intellectual disabilities. In addition, PCDH19 has been shown to play an important role in organizing cellular interactions in early neurodevelopmental processes. Ultimately, by gaining a greater understanding of the functions for PCDH19, we can further understand neurodevelopment and various neurodevelopmental diseases. For many in vitro studies of PCDH19, we first must obtain protein for use in the experiments. Human cell culture can be used to produces proteins of interest, when transfected with vectors containing the gene of interest. Here we tested two common transfection methods, lipofectamine transfection and calcium phosphate transfection, to determine which is the most cost effective method for the production of a PCDH19-GFP fusion protein. The cost per transfection is approximately five times higher per transfection with lipofectamine compared to calcium phosphate. However, the protein levels detected from lipofectamine were also consistently higher. The degree of increased protein production was variable and would require further replications to ascertain. From this data, we concluded that calcium phosphate is more cost effective, but lipofectamine may be necessary in some cases to reach sufficient levels of protein expression for some experiments.
