US 11,959,084 B2
Nucleic acid construct
Shaun Cordoba, London (GB); James Sillibourne, London (GB); and Martin Pulé, London (GB)
Assigned to AUTOLUS LIMITED, London (GB)
Appl. No. 17/254,001
Filed by AUTOLUS LIMITED, London (GB)
PCT Filed Jul. 31, 2019, PCT No. PCT/GB2019/052150
§ 371(c)(1), (2) Date Dec. 18, 2020,
PCT Pub. No. WO2020/025953, PCT Pub. Date Feb. 6, 2020.
Claims priority of application No. 1812474 (GB), filed on Jul. 31, 2018.
Prior Publication US 2022/0106602 A1, Apr. 7, 2022
Int. Cl. C12N 15/67 (2006.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01)
CPC C12N 15/67 (2013.01) [C07K 14/70503 (2013.01); C07K 14/7051 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01)] 21 Claims
 
1. A nucleic acid construct comprising:
a first nucleotide sequence of interest (NOI1), a translational readthrough motif (TRM), and a second nucleotide sequence of interest (NOI2), and
a nucleotide sequence encoding a cleavage site (CL), so that NOI1 and NOI2 are expressed as separate proteins,
wherein the (TRM) comprises the sequence STOP-CUAG or STOP-CAAUUA, in which “STOP” is a stop codon.
 
10. A nucleic acid construct comprising:
a first nucleotide sequence of interest (NOI1), a translational readthrough motif (TRM), and a second nucleotide sequence of interest (NOI2), and
a nucleotide sequence encoding a cleavage site (CL), so that NOI1 and NOI2 are expressed as separate proteins,
wherein NOI2 encodes a cytokine, chemokine or toxin.
 
15. A nucleic acid construct comprising: a first nucleotide sequence of interest (NOI1), a translational readthrough motif (TRM) and a second nucleotide sequence of interest (NOI2),
wherein the nucleic acid construct is capable of producing two products when expressed in a cell:
a) a first product encoded by NOI1 alone, and
b) a second product, encoded by NOI1 and NOI2, which is produced when translational readthrough occurs, and
wherein the first and/or second product is a chimeric antigen receptor (CAR).