Investigating the Relationship Between Masp1 and Bmp4 Protein Function Using Dual-Luciferase Assay
Additional Funding Sources
This project was made possible by the NSF Idaho EPSCoR Program and by the National Science Foundation under Award No. OIA-1757324.
Presentation Date
7-2022
Abstract
MASP1 is a serine protease known for its involvement in the innate immune system. Mutations in the gene that codes for MASP1 (MASP1/3) have been found in patients with 3MC Syndrome, a developmental disorder that causes craniofacial malformation, suggesting that Masp1 is important for embryonic development. However, the function of Masp1 during development is unknown. Using the African Clawed Frog (Xenopus laevis) as a model system, we previously found that genetic manipulation of Masp1 expression level alters cement gland shape and size.3 Multiple genes are involved in regulating cement gland formation, including Bmp4, and we found that Bmp4 gene expression is altered in Masp1 manipulated embryos. For this project, we want to determine how Masp1 impacts Bmp4 function. To address this question, we are performing a Dual-Luciferase assay to measure Bmp4 signaling activity in Xenopus laevis embryos. We are currently working to optimize assay conditions. We have determined optimal firefly luciferase plasmid concentrations and our assay protocol and found that we can detect Bmp4 signal. We are working on optimizing concentrations for the renilla luciferase. Once optimized, this will be an effective assay that can be used in Xenopus laevis to study Bmp signaling in relation to Masp1.
Investigating the Relationship Between Masp1 and Bmp4 Protein Function Using Dual-Luciferase Assay
MASP1 is a serine protease known for its involvement in the innate immune system. Mutations in the gene that codes for MASP1 (MASP1/3) have been found in patients with 3MC Syndrome, a developmental disorder that causes craniofacial malformation, suggesting that Masp1 is important for embryonic development. However, the function of Masp1 during development is unknown. Using the African Clawed Frog (Xenopus laevis) as a model system, we previously found that genetic manipulation of Masp1 expression level alters cement gland shape and size.3 Multiple genes are involved in regulating cement gland formation, including Bmp4, and we found that Bmp4 gene expression is altered in Masp1 manipulated embryos. For this project, we want to determine how Masp1 impacts Bmp4 function. To address this question, we are performing a Dual-Luciferase assay to measure Bmp4 signaling activity in Xenopus laevis embryos. We are currently working to optimize assay conditions. We have determined optimal firefly luciferase plasmid concentrations and our assay protocol and found that we can detect Bmp4 signal. We are working on optimizing concentrations for the renilla luciferase. Once optimized, this will be an effective assay that can be used in Xenopus laevis to study Bmp signaling in relation to Masp1.