IP Library › Granted Patent US 9,057,057
Granted Patent B2
US 9,057,057 · App. 13/812,857 · Granted Jun 16, 2015

Obligate heterodimer variants of foki cleavage domain

Inventors: Srinivasan Chandrasegaran (Baltimore, MD); Sivaprakash Ramalingam (Baltimore, MD); Karthikeyan Kandavelou (Pondicherry, IN)
Assignees: THE JOHNS HOPKINS UNIVERSITY; PONDICHERRY BIOTECH PRIVATE LIMITED (PBPL)
C12N9/16C07K14/4702C07K2319/70
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Quick Facts
Patent No.
US 9,057,057
App. No.
13/812,857
Granted
Jun 16, 2015
Kind
B2
Abstract

Disclosed are methods of making and using engineered FokI cleavage domain variants. Also disclosed are methods, compositions and fusion proteins containing obligate heterodimers of engineered FokI cleavage domain variants and DNA binding domains, such as zinc finger protein (ZFP) domains and transcription activator-like effector (TALE) domains.

Claims (6)

1. A polypeptide comprising either a first engineered FokI cleavage domain variant D483R:Q486E:I499L (SEQ ID NO: 1), or a second engineered FokI cleavage domain variant R487D:E490K:I538K (SEQ ID NO: 3), the first engineered FokI cleavage domain variant forming an obligate heterodimer with the second engineered FokI cleavage domain variant.

2. The polypeptide of claim 1 , wherein the obligate heterodimer comprises a first monomer containing the polypeptide designated D483R:Q486E:I499L (SEQ ID NO: 1), and a second monomer containing the polypeptide designated R487D:E490K:I538K (SEQ ID NO: 3).

3. The polypeptide of claim 1 , further comprising a DNA-binding domain.

4. The polypeptide of claim 3 , wherein the DNA-binding domain comprises zinc finger protein (ZFP) domain or transcription activator-like effector (TALE) domain.

5. A polynucleotide encoding the polypeptide of the claim 1 .

6. An isolated cell or cell line comprising the polypeptide or a polynucleotide encoding the polypeptide of the claim 1 .

Assignments (1)
CONFIRMATORY LICENSE Recorded Dec 12, 2017
From: JOHNS HOPKINS UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 044840/0569 →
Continuity (2)
Provisional Application 61368024 · Jul 27, 2010
Related Publication 20130130350A1 · May 23, 2013