Basil Hubbard

US Patent:

20120164670, Jun 28, 2012

Filed:

Jun 22, 2010

Appl. No.:

13/380108

Inventors:

Basil Hubbard - Boston MA, US
David A. Sinclair - Chestnut Hill MA, US

Assignee:

President and Fellows of Harvard College - Cambridge MA

International Classification:

C12Q 1/34
C12Q 1/48

US Classification:

435 15, 435 18

Abstract:

The invention relates to methods and kits for measuring enzyme activity, including enzymes that produce nicotinamide.

US Patent:

20210403887, Dec 30, 2021

Filed:

Jun 23, 2021

Appl. No.:

17/355735

Inventors:

- Cambridge MA, US
Basil Hubbard - Cambridge MA, US
Ahmed Hussein Badran - Cambridge MA, US

Assignee:

President and Fellows of Harvard College - Cambridge MA

International Classification:

C12N 9/22
C12N 15/62
C12N 15/85
C12N 15/90

Abstract:

Engineered transcriptional activator-like effectors (TALEs) are versatile tools for genome manipulation with applications in research and clinical contexts. One current drawback of TALEs is that the 5′ nucleotide of the target is specific for thymine (T). TALE domains with alternative 5′ nucleotide specificities could expand the scope of DNA target sequences that can be bound by TALEs. Another drawback of TALEs is their tendency to bind and cleave off-target sequence, which hampers their clinical application and renders applications requiring high-fidelity binding unfeasible. This disclosure provides methods and strategies for the continuous evolution of proteins comprising DNA-binding domains, e.g., TALE domains. In some aspects, this disclosure provides methods and strategies for evolving such proteins under positive selection for a desired DNA-binding activity and/or under negative selection against one or more undesired (e.g., off-target) DNA-binding activities. Some aspects of this disclosure provide engineered TALE domains and TALEs comprising such engineered domains, e.g., TALE nucleases (TALENs), TALE transcriptional activators, TALE transcriptional repressors, and TALE epigenetic modification enzymes, with altered 5′ nucleotide specificities of target sequences. Engineered TALEs that target ATM with greater specificity are also provided.