Col11a1 Regulates Chondrocyte Behavior During Chondrogenesis
Abstract
Little is known about the relationship between the extracellular matrix protein Collagen Type XIa1 (Col11a1) and chondrogenesis, the process of mesenchymal cells differentiating into chondrocytes. We hypothesize that decreasing the expression of Col11a1 inhibits chondrogenesis. Using the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR associated protein 9) method and RNAi transfection, the Col11a1 gene expression was decreased in chondrocytes. In order to verify Col11a1 was decreased, relative gene expression was determined using quantitative real-time PCR (polymerase chain reaction) and protein expression by western blotting. Upon successful completion of decreasing Col11a1, we expect to see altered gene and protein expression levels of chondrogenesis markers, such as Sox9, Col2a1, and Col10a1. We also expect to see a change in chondrocyte condensation and maturation as measured by proteoglycan accumulation. With these results, we understand Col11a1 is essential in establishing the cartilage architecture during chondrogenesis by modulating chondrocyte behavior.
Col11a1 Regulates Chondrocyte Behavior During Chondrogenesis
Little is known about the relationship between the extracellular matrix protein Collagen Type XIa1 (Col11a1) and chondrogenesis, the process of mesenchymal cells differentiating into chondrocytes. We hypothesize that decreasing the expression of Col11a1 inhibits chondrogenesis. Using the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR associated protein 9) method and RNAi transfection, the Col11a1 gene expression was decreased in chondrocytes. In order to verify Col11a1 was decreased, relative gene expression was determined using quantitative real-time PCR (polymerase chain reaction) and protein expression by western blotting. Upon successful completion of decreasing Col11a1, we expect to see altered gene and protein expression levels of chondrogenesis markers, such as Sox9, Col2a1, and Col10a1. We also expect to see a change in chondrocyte condensation and maturation as measured by proteoglycan accumulation. With these results, we understand Col11a1 is essential in establishing the cartilage architecture during chondrogenesis by modulating chondrocyte behavior.