US 11,884,925 B2
Synthetic immunomodulation with a CRISPR super-repressor in vivo
Samira Kiani, Scottsdale, AZ (US); Mo Reza Ebrahimkhani, Scottsdale, AZ (US); and Farzaneh Moghadam, Tempe, AZ (US)
Assigned to ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY, Scottsdale, AZ (US)
Appl. No. 17/292,407
Filed by ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY, Scottsdale, AZ (US)
PCT Filed Nov. 7, 2019, PCT No. PCT/US2019/060285
§ 371(c)(1), (2) Date May 7, 2021,
PCT Pub. No. WO2020/097344, PCT Pub. Date May 14, 2020.
Claims priority of provisional application 62/899,584, filed on Sep. 12, 2019.
Claims priority of provisional application 62/757,679, filed on Nov. 8, 2018.
Prior Publication US 2021/0380990 A1, Dec. 9, 2021
Int. Cl. C12N 15/63 (2006.01); C07K 14/47 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/86 (2006.01); A61K 48/00 (2006.01)
CPC C12N 15/635 (2013.01) [A61K 48/0008 (2013.01); C07K 14/4703 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/111 (2013.01); C12N 15/86 (2013.01); C07K 2319/85 (2013.01); C12N 2310/16 (2013.01); C12N 2310/20 (2017.05); C12N 2310/3519 (2013.01); C12N 2830/005 (2013.01)] 11 Claims
 
1. A synthetic repression system for repressing myeloid differentiation primary response 88 (MyD88) expression in vivo comprising:
(a) a nucleotide sequence encoding a guide RNA (gRNA) comprising
(i) a nucleic acid encoding a guide sequence comprising a sequence complementary to a portion of MyD88 and
(ii) an MS2 aptamer target site specific for an RNA binding bacteriophage MS2 coat protein;
(b) a nucleotide sequence encoding the RNA binding bacteriophage MS2 coat protein fused to a repression domain and comprising a sequence selected from the group consisting of SEQ ID NOs: 55, 59, 61, and 63; and
(c) a nucleotide sequence encoding a multifunctional Cas nuclease;
wherein the nucleotide sequences are packaged in at least one vector for viral delivery.