US 12,065,642 B2
Using nucleosome interacting protein domains to enhance targeted genome modification
Fuqiang Chen, St. Louis, MO (US); Xiao Ding, St. Louis, MO (US); Yongmei Feng, St. Louis, MO (US); and Gregory Davis, Webster Groves, MO (US)
Assigned to Sigma-Aldrich Co. LLC, St. Louis, MO (US)
Filed by Sigma-Aldrich Co. LLC, St. Louis, MO (US)
Filed on Feb. 18, 2020, as Appl. No. 16/793,272.
Application 16/793,272 is a continuation of application No. 16/031,819, filed on Jul. 10, 2018, granted, now 10,604,752.
Claims priority of provisional application 62/531,222, filed on Jul. 11, 2017.
Prior Publication US 2021/0024916 A1, Jan. 28, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01); C12N 15/85 (2006.01); C12N 15/90 (2006.01); C12Q 1/6897 (2018.01); A61K 48/00 (2006.01)
CPC C12N 15/102 (2013.01) [C12N 9/22 (2013.01); C12N 15/113 (2013.01); C12N 15/85 (2013.01); C12N 15/907 (2013.01); C12Q 1/6897 (2013.01); A61K 48/005 (2013.01); C07K 2319/09 (2013.01); C07K 2319/10 (2013.01); C07K 2319/80 (2013.01); C07K 2319/81 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 20 Claims
 
1. A fusion protein comprising a clustered regularly interspersed short palindromic repeats (CRISPR) protein directly or indirectly linked to two nucleosome interacting protein domains,
wherein the CRISPR protein is Streptococcus pyogenes Cas9 (SpCas9), Streptococcus pasteurianus Cas9 (SpaCas9), Francisella novicida Cpf1 (FnCpf1), or Campylobacter jejuni Cas9 (CjCas9),
wherein one of the two nucleosome interacting protein domains is a high mobility group (HMG) nucleosome-binding (HMGN) protein, and the other of the two nucleosome interacting protein domains is a high mobility group (HMG) box (HMGB) protein, a central globular domain from a histone H1 variant comprising SEQ ID NO:45, or a DNA binding domain from a chromatin remodeling complex protein, and
wherein the one of the two nucleosome interacting protein domains is directly or indirectly linked to the CRISPR protein at its N-terminus and the other of the two nucleosome interacting protein domains is directly or indirectly linked to the CRISPR protein at its C-terminus.