US 8,017,755 B2 · RNA-based transcriptional regulators

Abstract

RNA molecules have been shown to play a variety of functional roles in biological processes. RNA sequences can adopt 3-D conformations able to regulate transcription. RNA-based transcriptional regulators when recruited to a DNA template act to promote or suppress the transcription of nearby genes. The regulators are thought to act by mimicing the activation domains of protein transcriptional activators. These RNA-based transcriptional regulators may be engineered to regulate transcription based on the binding of a ligand such as a small molecule. RNA sequences of the regulators may be evolved to produce regulators with a greater degree of transcriptional activation or suppression. The RNA-based system of the present invention is useful in studying biological processes and in altering biological process in a therapeutic context.

Claims44

Dependent claims

An isolated transcriptional regulator comprising an RNA molecule, wherein the RNA molecule comprises a sequence having at least 80% homology with the sequence set forth as GGAUGCCUAAGCCUCUU (SEQ ID NO:

22), and wherein when the RNA molecule is recruited upstream of a gene, transcription of the gene is increased.

An isolated transcriptional regulator comprising an RNA molecule comprising a sequence selected from the group consisting of:

(SEQ ID NO:
23)
m26-10
CGCGGAAGAAUGAUCCCCCAAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
24)
m26-36
CGCGGAAGAAUUCUCCCCCAAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
25)
m26-31
CGCGGAAGAAUGCUCCCCCGAGUGGAUGCCUAAGCCUCAU;
(SEQ ID NO:
26)
m26-9
CGCGGAGGAAUGCUCCCCCGAGUGGAUGCCUAAUCCUCUU;
(SEQ ID NO:
27)
m26-14
CGCGGAAGCAUACUCCCCCGAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
28)
m26-15
CGCGGACCAAUGGUCCCCCAAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
29)
m26-34
CGCGGAAGAUUGCCCCCCCAAGUGGAUGCCUAAACCUCUA;
(SEQ ID NO:
30)
m26-29
CGCGGAAGAUUGUUCCCCCAAGUGGAUGCCUAAACCUCAU;
(SEQ ID NO:
31)
m26-28
CGCGGAAAAAUACUCCCCCAAGUGGAUGCCUAAACCUAUU;
(SEQ ID NO:
32)
m26-17
CGCGGAAGAAUGAUCCCCCAGGUGGAUGCCUAAGCCUCUA;
(SEQ ID NO:
33)
m26-30
CGCGGAACAAUGCUCCCCCAGCUGGAUGCCUAAGCCUCUA;
(SEQ ID NO:
34)
m26-20
CGGGGAAGAAUGCUGCCCAAAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
35)
m26-4
CGCGGAAGAAUGCUGCCCAACCUGGAUGCCUAAACCUCUU;
(SEQ ID NO:
36)
m26-32
CGCGGAAGAAUGCUGCCCAACCUGGAUGCCUAAACCUCUU;
(SEQ ID NO:
37)
m26-13
CUCGGAACAAUGCCCCCCCAAGUGGAUGCCUAAACCUCAU;
(SEQ ID NO:
38)
m26-35
CUCGGAAGAAUUCUCCCGAUAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
39)
m26-16
CGCGGAAGAAGAGUCCCCCGAGUGGAUGCCUAAACCUCUU;
(SEQ ID NO:
40)
m26-18
CGCGGAACAAUGUUGCCCCAAAUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
41)
m26-25
CGCGGAAGAACACUCCCCCACGUGGAUGCCUAAGCCUCCUU;
(SEQ ID NO:
42)
m26-21
CGCGGAAGACUGCAGCCCCAGGUGGAUGCCUAAACCUCGGU;
(SEQ ID NO:
43)
m26-39
CCCGGAAGACUGCAGCCCCAGGUGGAUGCCUAAACCUCUU;
(SEQ ID NO:
44)
m26-33
CGCGGAAGACUUCAGCCCCAAGUGGAUGCCUAAGCCUCUUA;
(SEQ ID NO:
45)
m26-1
CGCGGGUGUAAGCCCCCCCAGGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
46)
m26-23
CGCGUAAACAUGCGGCCCCAAGUGGAUGCCUAAGCCUCCU;
(SEQ ID NO:
47)
m26-24
CGCGGAAAGAUGAUCACCCGAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
48)
m26-11
CGCGCGAGUAUACUCCCCCAAGCGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
49)
m26-19
CGCAGAAGAUACCUCCCCCAAGUGGAUGCCUAAGCCUCAU;
(SEQ ID NO:
50)
m26-26
AACAGAAGAAUACUCCCCCAAGUGGAUACCUAAGCAUCUU;
(SEQ ID NO:
51)
m26-37
CGCGGAAAAAUUCUUCCCCCAAGUGGAUGCCUAAGCCUCUU;
(SEQ ID NO:
52)
m26-38
CGCGGAAAAAUUCUUCCCCCAAGUGGAUGCGUAAGCCUCUU;
(SEQ ID NO:
53)
m26-7
CGCGGAAGAACGCUCCCCGACGUGGAUGCCUAUUGUCCUU;
and
(SEQ ID NO:
54)
con-
CGCGGAAGAAUGCUCCCCCAAGUGGAUGCCUAAGCCUCUU.
sensus

127 more elements

Dependencies are inferred from the claim text.

Facts

Application

10/852,626Filed 24 May 2004 · us provisional application

Priority

23 May 2003

Published

US 2005/0070016 A131 Mar 2005

Granted

13 Sep 2011

Term ends

Not estimatedThe source records don’t support an estimate.

Classification10 codes

C12N 15/1086
Isolating an individual clone by screening libraries: preparation or screening of expression libraries, e.g. reporter assays
C12N 15/1048
Isolating an individual clone by screening libraries: selex
C12N 15/115
DNA or RNA fragments; Modified forms thereof ; Non-coding nucleic acids having a biological activity: aptamers, i.e. nucleic acids binding a…
C12N 15/63
Recombinant DNA-technology: introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

RNA-based transcriptional regulators

Life of this patent

RNA-based transcriptional regulators

Life of this patent

Abstract

Claims44