IP Library › Granted Patent US 12,630,820
Granted Patent B2
US 12,630,820 · App. 17/231,556 · Granted May 19, 2026

Compositions and methods for immunotherapy

Inventors: Amy Melissa Becker (Lexington, MA); Surbhi Goel (Winchester, MA); Sarah Beth Hesse (Arlington, MA); Troy Aaron Luster (Norfolk, MA); Birgit Schultes (Arlington, MA); Stephanie A. Yazinski (Lynnfield, MA); Pooja Vinay (St. Louis, MO)
Assignee: Intellia Therapeutics, Inc.
C12N15/111C07K14/7051C12N9/22C12N2310/20C12N2320/30
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,630,820
App. No.
17/231,556
Granted
May 19, 2026
Kind
B2
Abstract

Compositions and methods for editing, e.g., altering a DNA sequence, within the TRBC1, TRBC2 and/or TRAC genes are provided. Compositions and methods for immunotherapy are provided, for example.

Claims (38)

1 . A method of reducing the expression of an endogenous T cell receptor and inserting a heterologous immunological receptor at the TRAC locus, comprising delivering to a cell: (a) a TRAC guide RNA and a TRBC1/2 guide RNA; (b) an RNA-guided DNA binding agent, or a nucleic acid encoding an RNA-guided DNA binding agent; and (c) a nucleic acid encoding the heterologous immunological receptor,

wherein the TRAC guide RNA comprises:

i) a guide sequence that is SEQ ID NO: 90; or

ii) a guide sequence that is SEQ ID NO: 185; or

iii) a guide sequence that is SEQ ID NO: 214.

2 . The method of claim 1 , wherein the nucleic acid encoding the heterologous immunological receptor is flanked by sequences homologous to the TRAC locus.

3 . The method of claim 1 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 90.

4 . The method of claim 1 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 90 and the TRBC1/2 guide RNA comprises a guide sequence that is SEQ ID NO: 2.

5 . The method of claim 1 , wherein the heterologous immunological receptor is a heterologous T-cell receptor.

6 . The method of claim 5 , wherein the heterologous T-cell receptor recognizes a cancer antigen.

7 . The method of claim 6 , wherein the heterologous T-cell receptor is a WT1-specific T-cell receptor that recognizes WT1 or a fragment thereof.

8 . The method of claim 1 , wherein the heterologous immunological receptor is a chimeric antigen receptor.

9 . The method of claim 8 , wherein the chimeric antigen receptor recognizes a cancer antigen.

10 . The method of claim 1 , wherein the RNA-guided DNA binding agent is Cas9.

11 . The method of claim 1 , wherein the TRAC guide RNA and the TRBC1/2 guide RNA, the RNA-guided DNA binding agent or the nucleic acid encoding the RNA-guided DNA binding agent, and the nucleic acid encoding the heterologous immunological receptor are delivered to the cell via a vector, via transfection, via a lipid nanoparticle, or via microinjection.

12 . The method of claim 1 , wherein the TRAC guide RNA comprises SEQ ID NO: 186.

13 . The method of claim 1 , wherein the TRAC guide RNA comprises SEQ ID NO: 203.

14 . The method of claim 1 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 185.

15 . The method of claim 1 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 214.

16 . The method of claim 1 , wherein the TRBC1/2 guide RNA comprises a guide sequence that is SEQ ID NO: 2, a guide sequence that is 95% or 90% identical to SEQ ID NO: 2, or a guide sequence that is 18 or 19 contiguous nucleotides of SEQ ID NO: 2.

17 . A method of reducing the expression of an endogenous T cell receptor and inserting a heterologous immunological receptor at the TRAC locus, comprising delivering to a cell: (a) a TRAC guide RNA; (b) an RNA-guided DNA binding agent, or a nucleic acid encoding an RNA-guided DNA binding agent; and (c) a nucleic acid encoding the heterologous immunological receptor,

wherein the TRAC guide RNA comprises:

i) a guide sequence that is SEQ ID NO: 90; or

ii) a guide sequence that is SEQ ID NO: 185; or

iii) a guide sequence that is SEQ ID NO: 214.

18 . The method of claim 17 , wherein the nucleic acid encoding the heterologous immunological receptor is flanked by sequences homologous to the TRAC locus.

19 . The method of claim 17 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 90.

20 . The method of claim 17 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 185.

21 . The method of claim 17 , wherein the TRAC guide RNA comprises a guide sequence that is SEQ ID NO: 214.

22 . The method of claim 17 , wherein the TRAC guide RNA comprises SEQ ID NO: 186.

23 . The method of claim 17 , wherein the TRAC guide RNA comprises SEQ ID NO: 203.

24 . The method of claim 17 , wherein the heterologous immunological receptor is a heterologous T-cell receptor.

25 . The method of claim 24 , wherein the heterologous T-cell receptor recognizes a cancer antigen.

26 . The method of claim 25 , wherein the heterologous T-cell receptor is a WT1-specific T-cell receptor that recognizes WT1 or a fragment thereof.

27 . The method of claim 17 , wherein the heterologous immunological receptor is a chimeric antigen receptor.

28 . The method of claim 27 , wherein the chimeric antigen receptor recognizes a cancer antigen.

29 . The method of claim 17 , wherein the RNA-guided DNA binding agent is Cas9.

30 . The method of claim 17 , wherein the TRAC guide RNA, the RNA-guided DNA binding agent or the nucleic acid encoding the RNA-guided DNA binding agent, and the nucleic acid encoding the heterologous immunological receptor are delivered to the cell via a vector, via transfection, via a lipid nanoparticle, or via microinjection.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 23, 2021
From: BECKER, AMY MELISSA; GOEL, SURBHI; HESSE, SARAH BETH; LUSTER, TROY AARON; SCHULTES, BIRGIT; YAZINSKI, STEPHANIE A.; VINAY, POOJA
To: INTELLIA THERAPEUTICS, INC.
Reel/Frame 056963/0001 →
Continuity (4)
Continuation PCTUS2019056399 · Oct 15, 2019
Provisional Application 62747037 · Oct 17, 2018
Provisional Application 62746522 · Oct 16, 2018
Related Publication 20210340530A1 · Nov 4, 2021
References Cited (89)
US 5378825A · Cook et al. · 1995 [cited by applicant]
US 5585481A · Arnold, Jr. et al. · 1996 [cited by applicant]
US 8889356B2 · Zhang · 2014 [cited by applicant]
US 9023649B2 · Mali et al. · 2015 [cited by applicant]
US 9890393B2 · Duchateau · 2018 [cited by examiner]
US 10934336B2 · Zhao · 2021 [cited by examiner]
US 20140186958A1 · Zhang · 2014 [cited by applicant]
US 20150166980A1 · Liu et al. · 2015 [cited by applicant]
US 20160083449A1 · Schmitt et al. · 2016 [cited by applicant]
US 20160312198A1 · Joung et al. · 2016 [cited by applicant]
US 20160312199A1 · Joung et al. · 2016 [cited by applicant]
US 20170114334A1 · May et al. · 2017 [cited by applicant]
US 20170175128A1 · Welstead et al. · 2017 [cited by applicant]
US 20200216805A1 · Yuan et al. · 2020 [cited by applicant]
US 20210017249A1 · Sather et al. · 2021 [cited by applicant]
US 20210137978A1 · Maus · 2021 [cited by examiner]
CN 106191062A · 2016 [cited by applicant]
CN 107723275A · 2018 [cited by applicant]
CN 107746831A · 2018 [cited by applicant]
CN 107828730A · 2018 [cited by applicant]
CN 108138183A · 2018 [cited by applicant]
EP 3309248B1 · 2018 [cited by applicant]
EP 3686275A1 · 2020 [cited by applicant]
EP 3699268A1 · 2020 [cited by applicant]
JP 2016525888A · 2016 [cited by applicant]
JP 2017535261A · 2017 [cited by applicant]
WO 1993013121A1 · 1993 [cited by applicant]
WO 1995032305 · 1995 [cited by applicant]
WO 2014136086 · 2014 [cited by applicant]
WO 2014153470A2 · 2014 [cited by applicant]
WO 2014191128A1 · 2014 [cited by applicant]
WO 2015095340 · 2015 [cited by applicant]
WO 2015136001A1 · 2015 [cited by applicant]
WO 2015161276A2 · 2015 [cited by applicant]
WO 2016069282A1 · 2016 [cited by applicant]
WO 2016069283A1 · 2016 [cited by applicant]
WO 2016073964A1 · 2016 [cited by applicant]
WO 2016073966A1 · 2016 [cited by applicant]
WO 2016154596A1 · 2016 [cited by applicant]
WO 2016160721A1 · 2016 [cited by applicant]
WO 2016161273A1 · 2016 [cited by applicant]
WO 2017062451A1 · 2017 [cited by applicant]
WO 2017070429A1 · 2017 [cited by applicant]
WO 2017093969A1 · 2017 [cited by applicant]
WO 2017106528A2 · 2017 [cited by applicant]
WO 2017112944A1 · 2017 [cited by applicant]
WO 2017136794A1 · 2017 [cited by applicant]
WO 2017152015A1 · 2017 [cited by applicant]
WO 2017173054 · 2017 [cited by applicant]
WO 2017180989A2 · 2017 [cited by applicant]
WO 2017193107A2 · 2017 [cited by applicant]
WO 2018073393A2 · 2018 [cited by applicant]
WO 2018107028A1 · 2018 [cited by applicant]
WO 2018115887A1 · 2018 [cited by applicant]
WO 2018132479A1 · 2018 [cited by applicant]
WO 2018191490A1 · 2018 [cited by applicant]
WO 2018197492A1 · 2018 [cited by applicant]
WO 2019052577A1 · 2019 [cited by applicant]
WO 2019067992A1 · 2019 [cited by applicant]
WO 2019070541A1 · 2019 [cited by applicant]
WO 2019086007A1 · 2019 [cited by applicant]
WO 2019089610A1 · 2019 [cited by applicant]
WO 2019097305A2 · 2019 [cited by applicant]
WO 2019195491A1 · 2019 [cited by applicant]
WO 2019195492A1 · 2019 [cited by applicant]
Provasi, E., Genovese, P., Lombardo, A. et al. Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer. Nat Med 18, 807â815 (2012). (Year: 2012). [cited by examiner]
Jiangtao Ren, Xiaojun Liu, Chongyun Fang, Shuguang Jiang, Carl H. June, Yangbing Zhao; Multiplex Genome Editing to Generate Universal CAR T Cells Resistant to PD1 Inhibition. Clin Cancer Res May 1, 2017; 23 (9): 2255â22… [cited by examiner]
Clarke et al. Enhanced bacterial immunity and mammalian genome editing via RNA polymerase-mediated dislodging of Cas9 from double strand DNA breaks. Mol Cell. Jul. 5, 2018;71(1):42-55.e8. (Year: 2018). [cited by examiner]
Abbas, Yazan M et al. “Structure of human IFIT1 with capped RNA reveals adaptable mRNA binding and mechanisms for sensing N1 and N2 ribose 2′-O methylations.” Proceedings of the National Academy of Sciences of the Unite… [cited by applicant]
Adams et al., “The Biochemistry of the Nucleic Acids”, ed., 11th ed., 1992. [cited by applicant]
Cameron et al., “Mapping the genomic landscape of CRISPR-Cas9 cleavage”, Nature Methods. 6, 600-606; 2017. [cited by applicant]
Dunbar et al., “ANARCI: antigen receptor numbering and receptor classification,” Bioinformatics. Jan. 15, 2016; 32(2):298-300. [cited by applicant]
Eyquem, J. et al. “Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumor rejection” Nature, vol. 543, No. 7643 (Feb. 22, 2017) pp. 113-117. [cited by applicant]
Gilbert et al., “CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes,” Cell 154:442-51 (2013). [cited by applicant]
Guo, P X, and B Moss., “Interaction and mutual stabilization of the two subunits of vaccinia virus mRNA capping enzyme coexpressed in [cited by applicant]
International Search Report and Written Opinion issued in PCT.US2019/056399 dated Apr. 1, 2020. [cited by applicant]
Kabat et al., “Sequences of Proteins of Immunological Interest”, U.S. Dept. of Health and Human Services, Public Health Service, National Institutes of Health, 1991. [cited by applicant]
Katibah, George E et al. “Broad and adaptable RNA structure recognition by the human interferon-induced tetratricopeptide repeat protein IFIT5.” Proceedings of the National Academy of Sciences of the United States of Am… [cited by applicant]
Makarova et al., “An updated evolutionary classification of CRISPR-Cas systems” Nat Rev Microbiol, 13(11):722-36 (2015). [cited by applicant]
Makarova et al. “Evolution and classification of the CRISPR-Cas systems.” Nature reviews. Microbiology vol. 9,6 (2011): 467-77. [cited by applicant]
Mali et al., “CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering,” Nat. Biotechnol. 31:833-8 (2013). [cited by applicant]
Mao, X. and Shuman, S. (1994) “Intrinsic RNA (Guanine-7) Methyltransferase Activity of the Vaccinia Virus Capping Enzyme D1 Subunit Is Stimulated by the D12 Subunit”, J. Biol. Chem. 269, 24472-24479. [cited by applicant]
Perez-Pinera et al., “RNA-guided gene activation by CRISPR-Cas9-based transcription factors,” Nat. Methods 10:973-6 (2013). [cited by applicant]
Qi et al., “Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression,” Cell 152:1173-83 (2013). [cited by applicant]
Shmakov et al., “Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems” Molecular Cell, 60:385-397 (2015). [cited by applicant]
Stepinski et al., (2001) “Synthesis and properties of mRNAs containing the novel ‘anti-reverse’ cap analogs 7-methyl (3′-O-methyl)GpppG and 7-methyl(3′deoxy)GpppG,” RNA 7: 1486-1495. [cited by applicant]
Vester and Wengel, 2004, “LNA (locked nucleic acid): high-affinity targeting of complementary RNA and DNA” Biochemistry 43(42):13233-41. [cited by applicant]
Zetsche et al., “Cpf1 is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System,” Cell, (2015) 163, 3:759-771. [cited by applicant]
Georgiadis et al., “Long Terminal Repeat CRISPR-CAR-Coupled “Universal” T Cells Mediate Potent Anti-leukemic Effects,” Molecular Therapy, vol. 26, No. 5, pp. 1215-1227 (Mar. 6, 2018). [cited by applicant]