2021-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/654509In Taiwan, the total amount of biotechnology spent on the treatment of infertility exceeds NT$20 billion per year; however, in the absence of a comprehensive understanding and evaluation system, causes for infertility problem cannot be fully explained and cure, and as a consequence, techniques facilitate fertilization success cannot be effectively improved. Recent studies demonstrated that proteins within seminal plasma have various functions to promote pregnancy success. We previously identified, through the proteomic analysis, a high level sulfhydroxylase QSOX (quiescin Q6/sulfhydryl oxidases) protein present in the semen, and exhibits a inter-species agglutination of abnormal sperm (eg: rodent QSOX1 can recognize and agglutinates human or pig sperm), further analysis of the QSOX found the existence of two protein isoforms with complementary tissue distribution in the male reproductive tract. Moreover, these isoforms associate at specific, but distinct sperm membrnae surface of maturating sperm cells. Therefore, we hypothesize that the QSOX proteins may play important roles in the regulation of sperm maturation process and its physiological functions. This project will use the CRISPR/Cas9 gene editing technology and the Cre-LoxP system to construct whole-gene knockout and organ-specific transgenic mice, and conduct in-depth systematic research on the function of QSOX genes and proteins in the male reproductive tract. In addition, the use of the established QSOX KO cell line (QSOX1-/-, QSOX2-/-) and fluorescent-tagged QSOX cell line (eGFP-QSOX1, eRFP-QSOX2) for in vitro 2D polarized cell culture system, we can investigate the molecular basis of QSOX effect on sperm function and the underlysing regulatory mechanism. Due to current lack of literature information on QSOX, this project is expected to gain insight into the role of QSOX genes and proteins in the process of sperm maturation, and to explore the feasibility of QSOX gene editing or the stimulation and inhibition of QSOX protein secretion to directly or indirectly affect sperm quality that lead to better fertilization success. The experimental system used in this project will allow further applications on the study of epithelium-cell/gamete interactions in a broad range of organs, for example to study acidification of renal lumen, the molecular regulation of oocyte maturation and sperm-oocyte interaction from the oviduct epithelium.spermatozoa; seminal vesicle proteins; infertility; reproductive tract