IP Library Granted Patent US 10,344,271
Granted Patent B2
US 10,344,271 · App. 14/990,580 · Granted Jul 9, 2019

Modified zinc finger binding proteins

Inventors: Edward J. Rebar (Richmond, CA); Andrew Jamieson (Richmond, CA)
Assignee: Sangamo Therapeutics, Inc.
C12N9/22C07K14/4702C12N9/1007C12N15/8216C12N15/8217C12N15/8243A61K48/00C07K2319/00C07K2319/81C12Y301/00
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Quick Facts
Patent No.
US 10,344,271
App. No.
14/990,580
Granted
Jul 9, 2019
Kind
B2
Abstract

Disclosed herein are compositions and method comprising non-canonical (e.g., non-C2H2) zinc finger proteins.

Claims (27)

1. A plant comprising one or more plant cells in which a gamma tocopherol methyltransferase (gmt) gene is modified at a sequence as shown in any of SEQ ID NOs: 39, 43, 47, 51, 55, 59, 63, 67, 71, 75, 79, 83, 87, 91, 95, or 99, by insertion of a transgene that encodes a GMT protein, by transforming the one or more plant cells with a vector encoding a zinc finger nuclease comprising a pair of zinc finger proteins, each zinc finger protein comprising at least three zinc finger domains ordered finger 1 to finger 3 (F1 to F3), each zinc finger domain comprising a recognition helix region, wherein the zinc finger nuclease comprises a zinc finger protein comprising the following recognition helix regions:

(i) F1: RSDNLAR (SEQ ID NO:40); F2: DRSNLTR (SEQ ID NO:41); and F3: RSDALTR (SEQ ID NO:42), which zinc finger protein binds to a target site as shown in SEQ ID NO:39; or

(ii) F1: RSDHLAR (SEQ ID NO:44); F2: TSGNLVR (SEQ ID NO:45); and F3: RSDHLRE (SEQ ID NO:46), which zinc finger protein binds to a target site as shown in SEQ ID NO:43; or

(iii) F1: RSDALTR (SEQ ID NO:48); F2: RSDHLTT (SEQ ID NO:49); and F3: RSDHLTT (SEQ ID NO:50), which zinc finger protein binds to a target site as shown in SEQ ID NO:47; or

(iv) F1: QSSNLAR (SEQ ID NO:52); F2: RSDNLAR (SEQ ID NO:53); and F3: QSGNLTR (SEQ ID NO:54), which zinc finger protein binds to a target site as shown in SEQ ID NO:51; or

(v) F1: RSDHLAR (SEQ ID NO:56); F2: QSGNLAR (SEQ ID NO:57); and S25-US4 F3: RSDNLTR (SEQ ID NO:58), which zinc finger protein binds to a target site as shown in SEQ ID NO:55; or

(vi) F1: ERGTLAR (SEQ ID NO:60); F2: QSGSLTR (SEQ ID NO:61); and F3: RSDHLTT (SEQ ID NO:62), which zinc finger protein binds to a target site as shown in SEQ ID NO:59; or

(vii) F1: RSDHLTQ (SEQ ID NO:64); F2: QSGNLAR (SEQ ID NO:65); and F3: RSDHLSR (SEQ ID NO:66), which zinc finger protein binds to a target site as shown in SEQ ID NO:63; or

(viii) F1: QSSHLAR (SEQ ID NO:68); F2: RSDNLAR (SEQ ID NO:69); and F3: QSGNLAR (SEQ ID NO:70), which zinc finger protein binds to a target site as shown in SEQ ID NO:67; or

(ix) F1: QSSNLQR (SEQ ID NO:72); F2: RSDNALR (SEQ ID NO:73); and F3: RSDNLQR (SEQ ID NO:74), which zinc finger protein binds to a target site as shown in SEQ ID NO:71; or

(x) F1: RSDNALR (SEQ ID NO:76); F2: RSDNLAR (SEQ ID NO:77); and F3: RSDNLTR (SEQ ID NO:78), which zinc finger protein binds to a target site as shown in SEQ ID NO:75; or

(xi) F1: QSSDLR (SEQ ID NO:80); F2: RSDELQR (SEQ ID NO:81); and F3: RSDALTR (SEQ ID NO:82), which zinc finger protein binds to a target site as shown in SEQ ID NO:79; or

(xii) F1: QSSNLAR (SEQ ID NO:84); F2: QSGHLQR (SEQ ID NO:85); and F3: RSDHLTT (SEQ ID NO:86), which zinc finger protein binds to a target site as shown in SEQ ID NO:83; or S25-US4

(xiii) F1: QSSDLRR (SEQ ID NO:88); F2: QSGNLAR (SEQ ID NO:89); and F3: RSDNLTR (SEQ ID NO:90), which zinc finger protein binds to a target site as shown in SEQ ID NO:87; or

(xiv) F1: DRSHLTR (SEQ ID NO:92); F2: TSGHLTT (SEQ ID NO:93); and F3: QSSDLTR (SEQ ID NO:94), which zinc finger protein binds to a target site as shown in SEQ ID NO:91; or

(xvi) F1: QSSNLAR (SEQ ID NO:96); F2: RSDALSR (SEQ ID NO:97); and F3: QSGSLTR (SEQ ID NO:98), which zinc finger protein binds to a target site as shown in SEQ ID NO:95; or

(xvii) F1: QSSNLAR (SEQ ID NO:100); F2: RSDHLTT (SEQ ID NO:101); and F3: RSDALTR (SEQ ID NO:102), which zinc finger protein binds to a target site as shown in SEQ ID NO:99,

such that expression of GMT protein is increased within the plant.

2. A plant part from the plant of claim 1 , comprising the one or more modified plant cells.

3. The plant part of claim 2 , wherein the plant part is an endosperm, aleurone layer, embryo, scutellum, cotyledon, pericarp, stem, leaf, tuber, or a root.

4. The plant of claim 1 , wherein the plant is selected from the group consisting of wheat, maize, rice, millet, barley, tomato, apple, pear, strawberry, orange, alfalfa, carrot, potato, sugar beets, yam, lettuce, spinach, petunia, rose, chrysanthemum , pine, fir, spruce, sunflower, rape seed, and Arabidopsis.

5. The plant of claim 1 , wherein the zinc finger nuclease comprises a non-canonical zinc finger domain.

6. The plant of claim 1 , wherein the zinc finger nuclease is introduced into the plant cell as a polynucleotide.

7. A method for producing the plant of claim 1 , the method comprising introducing one or more zinc finger nucleases into a plant cell such that expression of GMT protein within the cell is increased.

8. The method claim 7 , wherein the altered GMT expression results in increased production of pharmaceutical compounds or industrial chemicals.

9. A method of modulating expression of GMT protein, comprising planting the plant of claim 1 , or a seed or plant part of the plant.

10. The method of claim 9 , wherein the plant is selected from the group consisting of wheat, maize, rice, millet, barley, tomato, apple, pear, strawberry, orange, alfalfa, carrot, potato, sugar beets, yam, lettuce, spinach, petunia, rose, chrysanthemum , pine, fir, spruce, sunflower, rape seed, and Arabidopsis.

Assignments (1)
CHANGE OF NAME Recorded Sep 25, 2018
From: SANGAMO BIOSCIENCES, INC.
To: SANGAMO THERAPEUTICS, INC.
Reel/Frame 047639/0400 →
Continuity (5)
Continuation 11486158 · Jul 13, 2006
Division 10055711 · Jan 22, 2002
Provisional Application 60290716 · May 11, 2001
Provisional Application 60263445 · Jan 22, 2001
Related Publication 20160222364A1 · Aug 4, 2016