Abstract
Primary ciliary dyskinesia (PCD) is a rare genetic disease affecting one in 16,000 children and results from structural or functional defects of motile cilia1. Lack of functioning cilia may result in reduced respiratory function, subfertility or infertility, and hydrocephalus due to a reduced ability to clear fluids and particles. A cilium consists of nine microtubules centered around a central pair complex1. Mutations in genes Spef2 (bgh), Cfap221 (nm1054), or Cfap54(Cfap54gt/gt), which encode proteins located in the central pair complex, result in PCD2,3,4. We previously utilized single-cell RNA sequencing to identify differentially expressed genes (DEGs) in early and late deuterosomal cells, as well as ciliated epithelial cells. The results showed a predicted decrease in Ift172 and Ift74, our genes of interest (GOIs), in nm1054 and Cfap54gt/gtmutant mice5. The hypothesis is that there is a decreased quantity of our GOIs in late deuterosomal cells. To validate this hypothesis, we analyzed RNA expression from tracheas using quantitative RNAscope.
Quantifying and Validating the Decrease of Ciliogenesis Genes in Mouse Models with PCD
Primary ciliary dyskinesia (PCD) is a rare genetic disease affecting one in 16,000 children and results from structural or functional defects of motile cilia1. Lack of functioning cilia may result in reduced respiratory function, subfertility or infertility, and hydrocephalus due to a reduced ability to clear fluids and particles. A cilium consists of nine microtubules centered around a central pair complex1. Mutations in genes Spef2 (bgh), Cfap221 (nm1054), or Cfap54(Cfap54gt/gt), which encode proteins located in the central pair complex, result in PCD2,3,4. We previously utilized single-cell RNA sequencing to identify differentially expressed genes (DEGs) in early and late deuterosomal cells, as well as ciliated epithelial cells. The results showed a predicted decrease in Ift172 and Ift74, our genes of interest (GOIs), in nm1054 and Cfap54gt/gtmutant mice5. The hypothesis is that there is a decreased quantity of our GOIs in late deuterosomal cells. To validate this hypothesis, we analyzed RNA expression from tracheas using quantitative RNAscope.