title: DT40 knock-out and knock-in studies determine the regions necessary and sufficient for transcription and epigenetic conversion of the chicken Ig-β gene.
abstract: 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.
authors.affiliation: Department of Life Science, College of Science, Rikkyo University, Toshima-ku, Tokyo 171-8501, Japan.
authors.forename: Kakeru
authors.initials: K
authors.lastname: Itaya
authors.forename: Kozue
authors.initials: K
authors.lastname: Chayahara
authors.forename: Takanori
authors.initials: T
authors.lastname: Hirai
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Minbuta
authors.forename: Takafumi
authors.initials: T
authors.lastname: Uchikawa
authors.forename: Tomoki
authors.initials: T
authors.lastname: Tanaka
authors.forename: Shinya
authors.initials: S
authors.lastname: Masaki
authors.forename: Kosuke
authors.initials: K
authors.lastname: Kuroda
authors.forename: Masao
authors.initials: M
authors.lastname: Ono
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D002645:Chickens / Q000235:genetics* / Q000276:immunology; D003850:Deoxyribonuclease I / Q000378:metabolism; D057890:Epigenomics; D055879:Gene Knock-In Techniques / Q000379:methods*; D055786:Gene Knockout Techniques / Q000379:methods*; D007136:Immunoglobulins / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D014158:Transcription, Genetic
chemical_list: D007136:Immunoglobulins; C038767:immunoglobulin B; D003850:Deoxyribonuclease I