Methods and compositions for treating Huntington's Disease
Disclosed herein are compositions comprising non-naturally occurring zinc finger domains, fusion proteins comprising these zinc finger domains, polynucleotides encoding these proteins, cells expressing these proteins and pharmaceutical compositions comprising these proteins or polynucleotides as well as methods of modifying an Htt gene using these compositions for treating or preventing Huntington's Disease.
1. An mRNA encoding a fusion protein comprising a transcriptional repression domain and a non-naturally occurring zinc finger protein comprising 5 or 6 zinc finger domains ordered F1 to F5 or F1 to F6, respectively, wherein the zinc finger protein is designated 33074, 30080, 30643, 30648, 30645, 30640, 30642, or 30646 and specifically binds to mutant Huntington's gene allele (mHtt) gene and comprises the recognition helix regions as follows:
ZFP
F1
F2
F3
33074
RSDNLSE
KRCNLRC
QSGDLTR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 60)
NO: 61)
NO: 18)
33080
QSGDLTR
QSGDLTR
RSDNLSE
(SEQ ID
(SEQ ID
(SEQ ID
NO: 18)
NO: 18)
NO: 60)
30643
RSDNLSE
KRCNLRC
QSSDLSR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 60)
NO: 61)
NO: 31)
30648
RSDNLSE
KRCNLRC
RSDNLSE
(SEQ ID
(SEQ ID
(SEQ ID
NO: 60)
NO: 61)
NO: 60)
30645
RSDNLSE
KRCNLRC
QSSDLSR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 60)
NO: 61)
NO: 31)
30640
QSSDLSR
QWSTRKR
QSSDLSR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 31)
NO: 63)
NO: 31)
30642
QSGDLTR
QSSDLSR
QWSTRKR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 18)
NO: 31)
NO: 63)
30646
RSDNLSE
KRCNLRC
QSGDLTR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 60)
NO: 61)
NO: 18)
ZFP
F4
F5
F6
33074
QSGDLTR
RSDNLSE
KRCNLRC
(SEQ ID
(SEQ ID
(SEQ ID
NO: 18)
NO: 60)
NO: 61)
33080
KRCNLRC
QSGDLTR
QSGDLTR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 61)
NO: 18)
NO: 18)
30643
QWSTRKR
QSSDLSR
QWSTRKR
(SEQ ID
(SEQ ID
(SEQ ID
NO: 63)
NO: 31)
NO: 63)
30648
KRCNLRC
RSDNLSE
KRCNLRC
(SEQ ID
(SEQ ID
(SEQ ID
NO: 61)
NO: 60)
NO: 61)
30645
QWSTRKR
QSGDLTR
NA
(SEQ ID
(SEQ ID
NO: 63)
NO: 18)
30640
QWSTRKR
QSGDLTR
NA
(SEQ ID
(SEQ ID
NO: 63)
NO: 18)
30642
QSSDLSR
QWSTRKR
NA
(SEQ ID
(SEQ ID
NO: 31)
NO: 63)
30646
QSSDLSR
QWSTRKR
NA
(SEQ ID
(SEQ ID
NO: 31)
NO: 63)
2. The mRNA of claim 1 , wherein the zinc finger protein binds to sequences within the CAG repeat region of the mHtt gene.
3. A host cell comprising one or more mRNAs of claim 1 .
4. A pharmaceutical composition comprising one or more mRNAs according to claim 1 .
5. A method of modifying expression of an mHtt gene in a cell, the method comprising administering to the cell one or more polynucleotides encoding one or more mRNAs according to claim 1 .
6. A method of treating Huntington's Disease, the method comprising administering one or more mRNAs according to claim 1 to a subject in need thereof.
7. A method of treating Huntington's Disease, the method comprising administering the pharmaceutical composition according to claim 4 to a subject in need thereof.
8. The method of claim 6 , wherein the one or more mRNAs are administered to the striatum of the subject.
9. The method of claim 7 , wherein the pharmaceutical composition is administered to the striatum of the subject.