The effects of RNAi to deplete C8orf37 in Paramecium tetraurelia
Abstract
Paramecium tetraurelia is a unicellular ciliated protozoan that is often used in scientific studies due to its ease of culture, simple structure, and genetic tractability. The swimming speed and direction of these single celled organisms can also provide insight on ion channel, pump, and cilia function. These organisms are dependent on cilia for mobility and feeding. C8orf37 is a gene that encodes a ciliary protein, which other studies have shown is implicated Bardet-Biedl Syndrome, a ciliopathy that affects photoreceptor cilia degradation. In humans, C8orf37 mutations have been linked to early macular degeneration, an ophthalmic condition that causes central vision loss. RNA interference (RNAi) is a scientific method used to prevent the mRNA from a targeted gene from being translated into a functional protein. The purpose of this study is to generate a RNAi construct that will deplete the C8orf37 gene and identify phenotype and gene expression discrepancies between the wild-type and experimental cells. In order to achieve this, a RNAi construct was designed and introduced to Paramecium tetraurelia via feeding. Then, swimming assays, EVOS photography, and immunostaining were performed to evaluate changes in cilia shape and function. As C8orf37 is essential for proper ciliary function in Paramecium tetraurelia, we expect its depletion via RNAi would result in impaired cilia function, such as structural abnormalities and altered swimming behavior.
Primary Faculty Mentor Name
Megan Valentine
Status
Undergraduate
Student College
College of Arts and Sciences
Program/Major
Biomedical Engineering
Primary Research Category
Life Sciences
The effects of RNAi to deplete C8orf37 in Paramecium tetraurelia
Paramecium tetraurelia is a unicellular ciliated protozoan that is often used in scientific studies due to its ease of culture, simple structure, and genetic tractability. The swimming speed and direction of these single celled organisms can also provide insight on ion channel, pump, and cilia function. These organisms are dependent on cilia for mobility and feeding. C8orf37 is a gene that encodes a ciliary protein, which other studies have shown is implicated Bardet-Biedl Syndrome, a ciliopathy that affects photoreceptor cilia degradation. In humans, C8orf37 mutations have been linked to early macular degeneration, an ophthalmic condition that causes central vision loss. RNA interference (RNAi) is a scientific method used to prevent the mRNA from a targeted gene from being translated into a functional protein. The purpose of this study is to generate a RNAi construct that will deplete the C8orf37 gene and identify phenotype and gene expression discrepancies between the wild-type and experimental cells. In order to achieve this, a RNAi construct was designed and introduced to Paramecium tetraurelia via feeding. Then, swimming assays, EVOS photography, and immunostaining were performed to evaluate changes in cilia shape and function. As C8orf37 is essential for proper ciliary function in Paramecium tetraurelia, we expect its depletion via RNAi would result in impaired cilia function, such as structural abnormalities and altered swimming behavior.
