Method for analyzing of human’s clock gene expression pattern using beard follicle cells

Atsushi Haraguchi, Masaki Takahashi,
Takuya Yoda, Masahito Hosokawa,
Kunpei Tanisawa, Mitsuru Higuchi,
Haruko Takeyama, ○Shigenobu Shibata.

 In mammals, we have circadian clock, which governs 24-hr based physiological function such as sleep-wake cycle. The mammalian circadian system is composed by several clock genes, such as Cry1, Per2, Bmal1, Clock, and Rev-erbα, and generates a rhythm of approximately 24 hour by comprised of transcriptional and translational feedback loops that control the expression of these clock genes (Buhr & Takahashi, 2013).

To estimate human’s clock genes expression pattern, not only a thorough examination of the rhythmic pattern but also a convenient and less invasive method are required. Method for analyzing of human’s clock genes expression pattern using beard and/or hair follicle cells was established, although this method has not been confirmed well by other research groups (Akashi et al., 2010). To confirm this method, we have been evaluating the relationship between clock gene expression and circadian rhythm disorders, such as delayed sleep phase syndrome, night eating syndrome, and social jet lag.

On the other hand, the amount of RNA has been difference from one person to the next. To establish the new analysis method, which is less invasive for the human subjects than the conventional method, we required the method to analyze few amount of RNA accurately. Then, we tried to establish a new method for measuring human clock genes expression pattern by applying technique of sequencing for a single cell, Bead-seq (Matsunaga et al., 2015).