US 12,404,513 B2
Single-vector Type I vectors
Virginia Martinez, Copenhagen Ø (DK); Ruben Vazquez-Uribe, Copenhagen Ø (DK); Adam Takos, Copenhagen Ø (DK); and Eric Van Der Helm, Copenhagen Ø (DK)
Assigned to SNIPR BIOME APS, Copenhagen Ø (DK)
Filed by SNIPR Biome ApS, Copenhagen Ø (DK)
Filed on Nov. 8, 2023, as Appl. No. 18/504,952.
Application 18/504,952 is a continuation of application No. 16/201,736, filed on Nov. 27, 2018, granted, now 11,851,663.
Claims priority of application No. 1816700 (GB), filed on Oct. 14, 2018; and application No. 1817509 (GB), filed on Oct. 27, 2018.
Prior Publication US 2024/0141365 A1, May 2, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/70 (2006.01); A61K 31/7088 (2006.01); A61K 38/46 (2006.01); A61P 3/04 (2006.01); A61P 31/04 (2006.01); C07K 14/33 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01)
CPC C12N 15/70 (2013.01) [A61K 31/7088 (2013.01); A61K 38/465 (2013.01); A61P 3/04 (2018.01); A61P 31/04 (2018.01); C07K 14/33 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01); C12N 2820/002 (2013.01); C12N 2820/007 (2013.01); C12N 2820/55 (2013.01); C12N 2830/005 (2013.01)] 27 Claims
 
1. A production strain bacterial cell comprising a nucleic acid vector for introduction into a target bacterial host cell for expression of Type I Cas3 and Cascade proteins in the target bacterial host cell, the vector comprising a first nucleotide sequence encoding a Type I Cas3 and a second nucleotide sequence encoding one or more cognate Cascade proteins, wherein the first nucleotide sequence is under the control of a promoter for controlling the expression of Type I Cas3 in the target bacterial host cell, wherein the promoter has a strength that is weaker than the Anderson Score strength of promoter BBa_J23108,
wherein the nucleic acid vector further comprises: (i) a CRISPR array for producing a crRNA in the target bacterial host cell; or (ii) a nucleotide sequence encoding a guide RNA (gRNA), wherein the crRNA or gRNA comprises a spacer sequence complementary to a target sequence of the target bacterial host cell, and
wherein the production strain bacterial cell does not comprise a crRNA or gRNA operable with the Cas3 to target and cut a chromosomal sequence of the production strain cell.