Abstract:
In recent days, many kinds of technologies including ChIP-sequencing, RNA-sequencing and
genome-editing using Crispr/Cas9 system have been applied to molecular biology, agricultural
sciences and so on. Especially genome editing technology is though to be powerful tool for
targeted mutageneeis in various organisms and this high mutagenesis efficiency have accelerated the use of this system in various kind of researches. Actually in agricultural field, this has
been applied to generate a tomato, rice and so on, which have desired characteristics. Today I
would like to show how we have used these new technologies to demonstrate the mechanism of
SOX9, a transcription factor, which regulates chondrogenesis. In vertebrae, most of the skeleton
is formed through endochondral ossification. Endochondral bone formation requires multiple
steps. SOX9 (SRY (sex-determining region Y)-related HMG box containing protein 9), a transcription factor, has been shown to contribute to the overt steps of chondrogenesis as a master
regulatory factor. Here I would like to show that many kinds of genes including ECM proteins,
ECM modification enzymes, receptors and transportors have been proven to be targets of this
transcription factor by use of RNA-seq and ChIP-seq. Also I would like to show this transcription factor regulates one of its most major target genes, Type II collagen α, Col2a1 through
its binding to an enhancer in its intron 6 by use of Cripsr/Cas9 mediated mutagenesis.
