title: Specific inhibition of class II MHC gene expression by anti-sense RNA.
abstract: We have established an anti-sense RNA system which is capable of regulating expression of the class II (Ia) molecule coded for by the major histocompatibility complex in cultured mouse cells. Various areas of the I-A beta chain gene were subcloned in an anti-sense orientation to the 3' of the dihydrofolate reductase (DHFR) cDNA under the control of the human metallothionein IIa gene promoter. These anti-sense DNA constructs were transfected into M12.4 cells, a BALB/c B lymphoma cell line which expresses both I-A and I-E molecules on the cell surface. I-A expression of selected clones transfected with anti-sense DNA encompassing the 5' untranslated region (UT) (100 or 310 bp) including the translation start site or the poly(A) addition signalling sequence in the 3' UT (250 bp) of the I-A beta chain gene were specifically reduced to less than 5% of the control M12.4 cell surface I-A expression. These clones had normal levels of I-E expression. However, transfection of the anti-sense DNA to the beta 1 domain (510 bp) including the splicing donor and acceptor sequences did not affect the expression of I-A molecules. The same antisense DNA constructs (100 bp of the 5' UT or 250 bp of the 3' UT) without the DHFR cDNA (710 bp) did not down-regulate the expression of I-A molecules, indicating that either the physical length of the anti-sense RNA or specific DHFR cDNA sequences are also important.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Immunology, ICMR, Kobe University School of Medicine, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Hatano
authors.forename: S
authors.initials: S
authors.lastname: Aizawa
authors.forename: T
authors.initials: T
authors.lastname: Soejima
authors.forename: K
authors.initials: K
authors.lastname: Shigemoto
authors.forename: M
authors.initials: M
authors.lastname: Taniguchi
authors.forename: T
authors.initials: T
authors.lastname: Tokuhisa
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005802:Genes, MHC Class II; D000949:Histocompatibility Antigens Class II / Q000235:genetics; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000276:immunology; D051379:Mice; D016372:RNA, Antisense / Q000235:genetics*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
chemical_list: D000949:Histocompatibility Antigens Class II; D016372:RNA, Antisense; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase