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Cholinergic Neuron‐Specific Expression of the Human Choline Acetyltransferase Gene Is Controlled by Silencer Elements

Abstract: Choline acetyltransferase (ChAT) is specifically expressed in Cholinergic neurons. To identify control mechanisms regulating the cell‐specific expression of the gene encoding ChAT, transient expression of the luciferase gene driven by human ChAT gene 5’ flanking sequences was compared in cholinergic and noncholinergic cell lines. Analysis of the gene indicated the presence of two regulatory elements with selective silencing activity. These elements, located between nucleotides −2043 to −3347 and nucleotides −3347 to −6550, act cooperatively to repress promoter activity > 10‐fold in a human adrenergic neuroblastoma cell line, SHSY5Y, and a human osteosarcoma cell line, 143 TK, while exhibiting less than a two‐fold effect in Cholinergic cell lines. Deletion of either nucleotides −2043 to −3347 or nucleotides −3348 to −6550 reduced cell‐specific repression by approximately half. Such differential repression appears to be responsible for the selective expression of the ChAT component of the Cholinergic phenotype.