US 12,146,880 B2
Artificial transcription factors comprising a sliding domain and uses thereof
Paul Blainey, Cambridge, MA (US); Anthony Kulesa, Cambridge, MA (US); and Kan Xiong, Cambridge, MA (US)
Assigned to The Broad Institute, Inc., Cambridge, MA (US); and Massachusetts Institute of Technology, Cambridge, MA (US)
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed on Oct. 13, 2020, as Appl. No. 17/069,558.
Application 17/069,558 is a continuation of application No. 15/099,371, filed on Apr. 14, 2016, granted, now 10,816,547.
Application 15/099,371 is a continuation in part of application No. PCT/US2014/060438, filed on Oct. 14, 2014.
Claims priority of provisional application 61/890,758, filed on Oct. 14, 2013.
Prior Publication US 2021/0300974 A1, Sep. 30, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/569 (2006.01); A61K 31/4375 (2006.01); A61K 47/54 (2017.01); A61K 47/64 (2017.01); C07K 14/00 (2006.01); C07K 14/39 (2006.01); C07K 14/47 (2006.01); C12N 15/67 (2006.01); C12Q 1/6848 (2018.01); C12Q 1/686 (2018.01)
CPC G01N 33/56966 (2013.01) [A61K 31/4375 (2013.01); A61K 47/549 (2017.08); A61K 47/64 (2017.08); A61K 47/645 (2017.08); C07K 14/00 (2013.01); C07K 14/39 (2013.01); C07K 14/4705 (2013.01); C12N 15/67 (2013.01); C12Q 1/6848 (2013.01); C12Q 1/686 (2013.01); C07K 2319/01 (2013.01); C07K 2319/80 (2013.01); G01N 2800/00 (2013.01)] 37 Claims
 
1. A method for improving the speed at which a sequence-specific DNA binding domain identifies a cognate DNA binding sequence in a target DNA molecule, wherein the sequence-specific DNA binding domain is contained within a non-naturally occurring or engineered artificial transcription factor polypeptide, the method comprising operably connecting via one or more linkers the sequence-specific DNA binding domain of the non-naturally occurring or engineered artificial transcription factor polypeptide with a sliding domain polypeptide, wherein (i) the sliding domain polypeptide comprises KRRR (SEQ ID NO: 2) and retains the ability to rapidly slide on DNA on DNA and (ii) the sequence-specific DNA binding domain comprises NVKRRTHNVLERQRRNE (SEQ ID NO: 21) or KRARNTEAARRSRAR (SEQ ID NO: 20),
thereby improving the speed at which the sequence-specific DNA binding domain of the non-naturally occurring or engineered artificial transcription factor identifies the cognate DNA binding sequence in the target DNA molecule, as compared to an appropriate control polypeptide composition lacking the sliding domain.