Inhibition of Expression of a Mouse .ALPHA.-Globin Gene by Plasmids that Include Antisense Oligonucleotides.

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Plasmid-borne DNAs, corresponding to 68-base oligodeoxynucleotides, synthesized in the anti-sense or sense configuration and based on the nucleotide sequences of various regions of the mouse α-globin mRNA, were introduced with the gene for xanthine-guanine phosphoribosyl transferase from E.coli(Ecogpt)into mouse erythroleukemia(MEL)cells by protoplast fusion.Specific inhibition of the synthesis of α-globin was observed only in the cells transformed with the plasmids with antisense 68-mers that corresponded to the cap site as well as the site of initiation of translation of α-globin mRNA(Oligo-A);Other plasmids with anti-sense 68-mers that corresponded to the regions of the exon/intron junctions, the individual exons, or the 3'un-translated region were ineffective.This antisense RNA efficiently reduced the production of α-globin to 9-18%of the endogenous level after induction with hexylmethylene-bis-acetoamide(HMBA).Moreover, most of the antisense transformants did not show any decrease in the expression of the c-myc gene at the early phases of differentiation of MEL cells.Thus, we propose a hypothesis that the early declineinlevels of c-myc mRNA may be independent of and uncoupled from the program of globin synthesis during the differentiation of MEL cells.
日本細胞生物学会の論文