Methods and compositions for treatment of a genetic condition

Patent metadata

Field	Value
Publication number	US-9963715-B2
Application number	US-201314013236-A
Country	US
Kind code	B2
Filing date	Aug 29, 2013
Priority date	Aug 29, 2012
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

Disclosed herein are nucleases and methods of using these nucleases for alteration of a globin gene and generation of cells.

First claim

What is claimed is:

1. A zinc finger protein comprising five or six zinc finger domains designated and ordered F1 to F5 or F1 to F6 from N-terminus to C-terminus, wherein F1 to F5 or F1 to F6 comprises any one of the following recognition helix regions:
(i) F1: DRSNLSR (SEQ ID NO:9);
F2: LRQNLIM (SEQ ID NO:161);
F3: TSANLTV (SEQ ID NO:162);
F4: RSDHLSR (SEQ ID NO:94);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QRNDRKS (SEQ ID NO:163), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:160;

(ii) F1: DRSNLSR (SEQ ID NO:9);
F2: LRQNLIM (SEQ ID NO:161);
F3: LQSQLNR (SEQ ID NO:164);
F4: RSDHLSR (SEQ ID NO:94);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QRNDRKS (SEQ ID NO:163), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:160;

(iii) F1: DRANLSR (SEQ ID NO:165);
F2: LRQNLIM (SEQ ID NO:161);
F3: LQSQLNR (SEQ ID NO:164);
F4: RSDHLSR (SEQ ID NO:94);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QRNDRKS (SEQ ID NO:163), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:160;

(iv) F1: DRSNLSR (SEQ ID NO:9);
F2: TSSNRNH (SEQ ID NO:166);
F3: HSGNLTK (SEQ ID NO:167);
F4: RSDHLSR (SEQ ID NO:94);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QKVDLSR (SEQ ID NO:168), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:160;

(v) F1: DRSNLSR (SEQ ID NO:9);
F2: TSSNRNH (SEQ ID NO:166);
F3: QANNLKV (SEQ ID NO:169);
F4: RSDHLSR (SEQ ID NO:94);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QKVDLSR (SEQ ID NO:168), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:160;

(vi) F1: RSDHLSA (SEQ ID NO:59);
F2: DRSALAR (SEQ ID NO:30);
F3: RSDSLSR (SEQ ID NO:171);
F4: DRSVRTK (SEQ ID NO:172);
F5: RSDHLSA (SEQ ID NO:59); and
F6: QRSNLKV (SEQ ID NO:173), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:170;

(vii) F1: RSDHLTQ (SEQ ID NO:35);
F2: DRSALAR (SEQ ID NO:30);
F3: RSDSLSR (SEQ ID NO:171);
F4: DRSVRTK (SEQ ID NO:172);
F5: RSDHLSA (SEQ ID NO:59); and
F6: QRSNLKV (SEQ ID NO:173), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:170;

(viii) F1: RSDHLTT (SEQ ID NO:174);
F2: DRSALAR (SEQ ID NO:30);
F3: RSDSLSR (SEQ ID NO:171);
F4: DRSVRTK (SEQ ID NO:172);
F5: RSDHLSA (SEQ ID NO:59); and
F6: QRSNLKV (SEQ ID NO:173), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:170;

(ix) F1: RSDHLSA (SEQ ID NO:59);
F2: WATARDR (SEQ ID NO:175);
F3: RSDSLSR (SEQ ID NO:171);
F4: HTKSLSR (SEQ ID NO:176);
F5: RSDHLSA (SEQ ID NO:59);
F6: QRSNLKV (SEQ ID NO:173); wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:170;

(x) F1: RSAHLTQ (SEQ ID NO:177);
F2: DRSVLRR (SEQ ID NO:178)
F3: RSDSLSR (SEQ ID NO:171);
F4: DRSVRTK (SEQ ID NO:172);
F5: RSDHLSA (SEQ ID NO:59); and
F6: QRSNLKV (SEQ ID NO:173), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:170;

(xi) F1: RSDVLSE (SEQ ID NO:57);
F2: RNQHRKT (SEQ ID NO:58);
F3: RSDHLSA (SEQ ID NO:59);
F4: RSANLTR (SEQ ID NO:60;
F5: RSDVLSN (SEQ ID NO:61); and
F6: DRSTRIT (SEQ ID NO:62), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:56;

(xii) F1: DRSNLSR (SEQ ID NO:9);
F2: HRQHLVT (SEQ ID NO:64);
F3: DRSNLTR (SEQ ID NO:65);
F4: QSGDLTR (SEQ ID NO:13); and
F5: HRSSLLN (SEQ ID NO:66), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:63;

(xiii) F1: QSGHLSR (SEQ ID NO:32);
F2: RSDHLST (SEQ ID NO:67);
F3: RSADLSR (SEQ ID NO:68);
F4: RSDNLSQ (SEQ ID NO:69); and
F5: ASNDRKK (SEQ ID NO:70), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:66; or

(xiv) F1: RSDNLSA (SEQ ID NO:72);
F2: RNNDRKT (SEQ ID NO:73);
F3: DRSDLSR (SEQ ID NO:29);
F4: TSSNRTK (SEQ ID NO:74);
F5: QSGNLAR (SEQ ID NO:39); and
F6: QSGDLTR (SEQ ID NO:13), wherein the zinc finger protein binds to a target site as set forth in SEQ ID NO:71.

2. A fusion protein comprising the zinc finger protein of claim 1 and a cleavage domain or cleavage half-domain.

3. An isolated polynucleotide encoding the zinc finger proteins of claim 1 .

4. An isolated cell comprising the zinc finger proteins of claim 1 .

5. An isolated cell comprising the polynucleotides of claim 3 .

6. The cell of claim 5 , wherein the cell is a red blood cell precursor cell.

7. The cell of claim 5 , wherein the cell is a red blood cell (RBC).

8. The cell of claim 6 , wherein the precursor cell is a CD34+ hematopoietic stem cell.

9. A kit comprising the protein of claim 1 .

10. A kit comprising the polynucleotide of claim 3 .

11. An isolated cell comprising the fusion protein of claim 2 .

Assignees

• Sangamo Biosciences Inc
• Sangamo Therapeutics Inc

Inventors

• Cost Gregory J
• Gregory Philip D
• Guschin Dmitry
• Holmes Michael C
• Miller Jeffrey C
• Paschon David
• Rebar Edward J
• Reik Andreas
• Urnov Fyodor
• Zhang Lei

Classifications

• A61P7/06

  Antianaemics · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C07K14/805

  Haemoglobins; Myoglobins · CPC title

• C12N9/22 Primary

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• C07K14/4702

  Regulators; Modulating activity · CPC title