Genomic DNA fragments containing regulatory and coding sequences for the B2-subunit of the neuronal nicotinic acetylcholine receptor and transgenic animals made using these fragments or mutated fragments

Abstract

Several genes encoding subunits of the neuronal nicotinic acetylcholine receptors have been cloned and regulatory elements involved in the transcription of the ∝:2 and ∝:7-subunit genes have been described. Yet, the detailed mechanisms governing the neuron-specific transcription and the spatio-temporal expression pattern of these genes remain largely uninvestigated. The β2-subunit is the most widely expressed neuronal nicotinic receptors subunit in the nervous system. We have studied the structural and regulatory properties of the 5′ sequence of this gene. A fragment of 1163 bp of upstream sequence is sufficient to drive the cell-specific transcription of a reporter gene in both transient transfection assays and in transgenic mice. Deletion analysis and site-directed mutagenesis of this promoter reveal two negative and one positive element. The positively acting sequence includes one functional E-box. One of the repressor elements is located in the transcribed region and is the NRSE / RE1 sequence already described in promoters of neuronal genes.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This is a continuation of application Ser. No. 10 / 843,432, filed May 12, 2004, which issued as U.S. Pat. No. 7,294,463 on Nov. 13, 2007, which is a continuation of application Ser. No. 09 / 349,925, filed Jul. 8, 1999, which issued as U.S. Pat. No. 6,777,236 on Aug. 17, 2004, which is a continuation of application Ser. No. 08 / 465,712, filed Jun. 6, 1995, which issued as U.S. Pat. No. 6,452,066 on Sep. 17, 2002, which is a continuation of application Ser. No. 08 / 358,627, filed Dec. 14, 1994, which issued as U.S. Pat. No. 6,177,242 on Jan. 23, 2001, all of which are incorporated herein by reference.BACKGROUND OF THE INVENTIONField of the Invention[0002]This invention relates to DNA and clones of β2-subunit of neuronal nicotinic acetylcholine receptor (nAChR) sequences. This invention also relates to genomic DNA fragments containing regulatory and coding sequences for the β2-subunit neuronal nAChR and transgenic animals made using these fragme...

AI Technical Summary

Benefits of technology

The patent text describes a gene structure and promoter region of the mouse nAchR β2-subunit gene. The researchers found that the promoter region is conserved among different mammalian species and contains many consensus motifs for nuclear protein binding. The promoter region is located between the Eco47III and BamHI sites and contains a NRSE / RE1 sequence. The researchers also tested the promoter in transgenic mice and found that it directs cell-specific expression of the nAchR β2-subunit gene. The promoter sequence can be used to control the expression of genes in neurons.

Problems solved by technology

Only limited information is available, to date, about the genetic mechanisms that account for regulation of nAchR gene transcription in neurons.

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