돼지 체세포 복제란 초기발달 과정 중 Dnmt1o상류 영역의 다이내믹한 DNA 메틸화 변화

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 36권 / 1호
ㆍ저자명 : 고응규, 김성우, 조상래, 도윤정, 김재환, 김상우, 김현, 박재홍, 박수봉

목차

ABSTRACT 서론 재료 및 방법 난자의 회수와 체외성숙 Donor Cell의 준비 체세포 복제란의 생산 체내수정란의 회수 Bisulfite Sequencing에 의한 DNA 메틸화 해석 Pyrosequencing에 의한 DNA 메틸화 해석 결과 체세포 복제란 초기 배 발달 중 Dnmt1o 상류 영역의 DNA메틸화 변화 체세포 복제란 초기 배 발달 중 Dnmt1s 상류 영역의 DNA메틸화 변화 Bisulfite Sequencing 방법에 의한 DNA 메틸화 분석 고찰 결론 인용문헌

영어 초록

DNA methyltransferase 1 (Dnmt1) gene contains three different isoform transcripts, Dnmt1s, Dnmt1o, and Dnmt1p, are produced by alternative usage of multiple first exons. Dnmt1o is specific to oocytes and preimplantation embryos, whereas Dnmt1s is expressed in somatic cells. Here we determined that porcine Dnmt1o gene had differentially methylated regions (DMRs) in 5’-flanking region, while those were not found in the Dnmt1s promoter region. The methylation patterns of the porcine Dnmt1o/Dnmt1s DMRs were investigated using bisulfite sequencing and pyrosequencing analysis through all preimplantation stages from one cell to blastocyst stage in in vivo or somatic cell nuclear transfer (SCNT). The Dnmt1o DMRs contained 8 CpG sites, which located in —640 bp to —30 bp upstream region from transcription start site of the Dnmt1o gene. The methylation status of 5 CpGs within the Dnmt1o DMRs were distinctively different at each stage from one-cell to blastocyst stage in the in vivo or SCNT, respectively. 55.62% methylation degree of the Dnmt1o DMRs in the in vivo was increased up to 84.38% in the SCNT embryo, moreover, de novo methylation and demethylation occurred during development of porcine embryos from the one-cell stage to the blastocyst stage. However, the DNA methylation states at CpG sites in the Dnmt1s promoter regions were hypomethylated, and dramatically not changed through one-cell to blastocyst stage in the in vivo or SCNT embryos. In the present study, we demonstrated that the DMRs in the promoter region of the porcine Dnmt1o was well conserved, contributing to establishment and maintenance of genome-wide patterns of DNA methylation in early embryonic development.

참고 자료

없음