US 12,419,955 B2
Method for efficiently infecting human natural killer cells and other immune cells with pseudovirus
Yongfeng Liu, Beijing (CN); and Zhihua Lu, Beijing (CN)
Assigned to Beijing Gene MINK Biotechnology CO.LTD, Beijing (CN)
Filed by Beijing Gene MINK Biotechnology CO.LTD, Beijing (CN)
Filed on Sep. 30, 2024, as Appl. No. 18/900,968.
Application 18/900,968 is a continuation of application No. PCT/CN2023/112838, filed on Aug. 14, 2023.
Claims priority of application No. 202310476442.2 (CN), filed on Apr. 28, 2023; and application No. 202310489784.8 (CN), filed on May 4, 2023.
Prior Publication US 2025/0018035 A1, Jan. 16, 2025
Int. Cl. A61K 39/00 (2006.01); A61K 40/11 (2025.01); A61K 40/15 (2025.01); A61K 40/31 (2025.01); A61K 40/42 (2025.01); C07K 14/725 (2006.01); C07K 16/28 (2006.01); C12N 15/86 (2006.01)
CPC A61K 40/4202 (2025.01) [A61K 40/11 (2025.01); A61K 40/15 (2025.01); A61K 40/31 (2025.01); C07K 14/7051 (2013.01); C07K 16/2878 (2013.01); C12N 15/86 (2013.01); C12N 2710/16143 (2013.01)] 7 Claims
 
1. A method for preparing natural killer (NK) cells expressing a target gene, comprising the steps of transfecting cells with a vector composition containing an envelope plasmid, collecting a viral liquid, and contacting the viral liquid with NK cells,
the envelope plasmid encoding a fusion protein having an X-Y-Z structure, the X having an amino acid sequence as shown in SEQ ID NO. 1, the Y having an amino acid sequence as shown in SEQ ID NO.2, and the Z having an amino acid sequence as shown in SEQ ID NO.3, wherein the envelope plasmid is configured to cause expression of the fusion protein in the transfected cells,
wherein the vector composition further contains the following plasmids:
1) An expression plasmid containing a sequence encoding a target gene, wherein the expression plasmid is configured to cause expression of the target gene in the transfected cells,
2) a packaging plasmid 1 containing a sequence encoding specific antigen (GAG), or polymerase (POL), or a combination thereof, wherein the expression plasmid is configured to cause expression said GAG, said POL, or said combination thereof in the transfected cells; and
3) A packaging plasmid 2 containing a sequence encoding a regulator of expression of virion protein (REV), wherein the expression plasmid is configured to cause expression of the REV in the transfected cells; and
wherein the fusion protein is QMV, which forms a pseudovirus named QMV virus targeting a receptor SLC20A1.