US 12,258,560 B2
Compositions and methods for transient gene therapy with enhanced stability
Michael Goldberg, Brookline, MA (US); and Ellese Carmona, Brookline, MA (US)
Assigned to DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US)
Filed by Dana-Farber Cancer Institute, Inc., Boston, MA (US)
Filed on Apr. 8, 2021, as Appl. No. 17/225,823.
Application 17/225,823 is a division of application No. 15/579,322, granted, now 11,000,547, previously published as PCT/US2016/036045, filed on Jun. 6, 2016.
Claims priority of provisional application 62/303,116, filed on Mar. 3, 2016.
Claims priority of provisional application 62/171,538, filed on Jun. 5, 2015.
Prior Publication US 2021/0299280 A1, Sep. 30, 2021
Int. Cl. C12N 15/11 (2006.01); A61K 35/17 (2015.01); A61K 48/00 (2006.01); A61P 35/00 (2006.01); C07K 14/005 (2006.01); C07K 14/195 (2006.01); C07K 14/435 (2006.01); C12N 5/0783 (2010.01)
CPC C12N 15/111 (2013.01) [A61K 35/17 (2013.01); A61K 48/0075 (2013.01); A61K 48/0091 (2013.01); A61P 35/00 (2018.01); C07K 14/005 (2013.01); C07K 14/195 (2013.01); C07K 14/435 (2013.01); C12N 5/0636 (2013.01); C12N 2320/51 (2013.01)] 16 Claims
 
1. A method for producing a chimeric antigen receptor (CAR) cell comprising contacting an immune cell with a circularized nucleic acid, wherein the circularized nucleic acid comprises:
a 5′ complement-reverse complement (CRC) sequence;
an internal ribosome entry site (IRES);
an RNA sequence encoding a chimeric antigen receptor (CAR);
a 3′ UTR sequence; and
a 3′ CRC sequence;
wherein the 5′ CRC sequence is fully complementary to the 3′ CRC sequence.