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US5215910A Expired / Lapsed US ⏱️ 0.0 years remaining

Host cells transformed with sterol regulatory elements

Assignee: The University of Texas System
•
Filing: 1990-06-01
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Granted: 1993-06-01
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Expires: 2010-06-01
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Abstract

Abstract Disclosed are discreet functionally translocatable DNA segments, termed Sterol Regulatory Elements (SRE's), which are fused to heterologous structural genes to provide sterol regulatory capability to the thus formed hybrid gene. The hybrid genes respond to sterols by decreasing the production of messenger RNA. The SRE segments contain as their primary functional nucleotide sequence, a 16 bp sequence referred to as direct repeat 2, isolated from the 5' regions of the human LDL receptor gene. DNA segments which include this 16 nucleotide long sequence similarly confer sterol regulatory capability to previously known promoters such as the HSV TK promoter. Also disclosed are discreet sequences which confer positive transcription promotion to heterologous structural genes and promoters without conferring sterol responsivity. Methods are disclosed for employing these genetic control elements in a myriad of embodiments which provide for a fine-tune control of heterologous genes. Methods are also disclosed for employing the SRE in a screening assay for drugs capable of stimulating the cell to synthesize LDL receptors.

Inventors

  • 👤 Michael S. Brown
  • 👤 Joseph L. Goldstein
  • 👤 David W. Russell
  • 👤 Thomas C. Sudhof

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-09 18:22
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85

Claims Analysis

Total Claims: 9
Claim 1
Independent Claim 29 words
A recombinant host cell transformed with a DNA containing a sterol regulatory element, wherein said sterol regulatory element comprises a nucleic acid sequence of: (a) 5'-A-A-A-A-T-C-A-C-C-C-C-A-C-T-G-C-3' or (b) 5'-G-C-A-G-T-G-G-G-G-T-G-A-T-T-T-T-3'.
Claim 2
Dependent Claim 29 words
A recombinant host cell transformed with a DNA containing a sterol regulatory element, wherein said sterol regulatory element comprises a nucleic acid sequence of: (a) 5'-A-A-A-A-T-C-A-C-C-C-C-A-C-T-G-C-3' or (b) 5'-G-C-A-G-T-G-G-G-G-T-G-A-T-T-T-T-3'.
Claim 3
Dependent Claim 21 words
The host cell of claim 1, wherein the sterol regulatory element is located on a recombinant DNA vector within the cell.
Claim 4
Dependent Claim 15 words
The host cell of claim 1, wherein the host cell is a eukaryotic host cell.
Claim 5
Dependent Claim 16 words
The host cell of claim 3, wherein the eukaryotic host cell comprises a mammalian host cell.
Claim 6
Dependent Claim 25 words
The host cell of claim 4, wherein the mammalian host cell comprises a Hep G2, VERO, HeLa, CHO, WI38, BHK, COS-7 or MDCK host cell.
Claim 7
Dependent Claim 87 words
The host cell of claim 1, further comprising a positive LDL receptor gene promoter element in operable linkage with said sterol regulatory element, wherein said positive LDL receptor gene promoter element comprises a nucleic acid sequence of: X n , wherein n=2-5, each X being independently selected from the DNA having the nucleotide sequence of: a) 5'-A-A-A-C-T-C-C-T-C-C-T-C-T-T-G-C-3'; b) 5'-A-A-A-C-T-C-C-T-C-C-C-C-C-T-G-C-3'; c) 5'-G-C-A-A-G-A-G-G-A-G-G-A-G-T-T-T-3'; or d) 5'-G-C-A-G-G-G-G-G-A-G-G-A-G-T-T-T-3'; with each X unit being separated by from 0-20 nucleotides selected from the group of nucleotides consisting of A, G, C, and T.
Claim 8
Dependent Claim 33 words
The host cell of claim 1, wherein the sterol regulatory element is positioned adjacent to a selected structural gene such that expression of the gene can be controlled by the sterol regulatory element.
Claim 9
Dependent Claim 14 words
The host cell of claim 7, wherein the structural gene comprises a reporter gene.

References Cited (Backward Citations)

WO1983001783A1 Cited
US4738922A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1996-05-31 1996-11-30 1997-06-01 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2000-05-31 2000-11-30 2001-06-01 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2004-05-31 2004-11-30 2005-06-01 $7700.00 Window Closed / Lapsed

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