Inhibition of expression of a mouse alpha-globin gene by plasmids that include antisense oligonucleotides.
Plasmid-borne DNAs, corresponding to 68-base oligodeoxynucleotides, synthesized in the antisense or sense configuration and based on the nucleotide sequences of various regions of the mouse alpha-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 alpha-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 alpha-globin mRNA (Oligo-A); Other plasmids with antisense 68-mers that corresponded to the regions of the exon/intron junctions, the individual exons, or the 3' untranslated region were ineffective. This antisense RNA efficiently reduced the production of alpha-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 decline in levels of c-myc mRNA may be independent of and uncoupled from the program of globin synthesis during the differentiation of MEL cells.
Yokoyama
K
K
Gene Bank, Tsukuba Life Science Center, RIKEN (The Institute of Physical and Chemical Research), Ibaraki, Japan.
Hou
D X
DX
Gao
H
H
Tang
X
X
Kitabayashi
I
I
Nishikura
K
K
Gachelin
G
G
D000081:Acetamides / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D004274:DNA, Recombinant / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D016259:Genes, myc; D005914:Globins / Q000096:biosynthesis* / Q000235:genetics; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010957:Plasmids; D012315:RNA Caps / Q000235:genetics; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D014407:Tumor Cells, Cultured
D000081:Acetamides; D004274:DNA, Recombinant; D012315:RNA Caps; D016372:RNA, Antisense; D012333:RNA, Messenger; D005914:Globins; C014026:hexamethylene bisacetamide