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DT40 knock‐out and knock‐in studies determine the regions necessary and sufficient for transcription and epigenetic conversion of the chicken Ig‐β gene
Author(s) -
Itaya Kakeru,
Chayahara Kozue,
Hirai Takanori,
Minbuta Tomohiro,
Uchikawa Takafumi,
Tanaka Tomoki,
Masaki Shinya,
Kuroda Kosuke,
Ono Masao
Publication year - 2011
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2011.01486.x
Subject(s) - biology , epigenetics , gene , intron , genetics , locus (genetics) , histone , regulation of gene expression , gene knockin , transcription (linguistics) , microbiology and biotechnology , dna methylation , hypersensitive site , gene expression , promoter , linguistics , philosophy
The chicken Ig‐β locus is organized by three cell‐type‐specific genes and two ubiquitously expressed genes. B‐cell‐specific DNase I hypersensitive sites (DHS) in that locus, including three present inside the flanking gene, were grouped into six regions and deleted. The deletions decreased Ig‐β mRNA content to <0.1% of that of normal DT40 cells and converted epigenetic parameters such as histone modifications, CG methylation and DNase I hypersensitivity into inactive states. Knocked‐in DHS regions into knock‐out cells reactivated both transcription of the Ig‐β gene and epigenetic parameters. Thus, the collaboration of the scattered regulatory regions was essential and sufficient not only for B‐cell‐specific transcription of the Ig‐β gene, but also for the conversion of epigenetic parameters. On the basis of the knock‐in studies, we determined the regions involved in the conversion and maintenance of the epigenetic parameters. These scattered regulatory regions were limited in vicinity such as in an intron of the gene, in the intergenic regions and in the introns of a flanking gene.