Targeted disruption of T cell receptor genes using engineered zinc finger protein nucleases

Patent metadata

Field	Value
Publication number	US-10918668-B2
Application number	US-201815936210-A
Country	US
Kind code	B2
Filing date	Mar 26, 2018
Priority date	Mar 21, 2013
Publication date	Feb 16, 2021
Grant date	Feb 16, 2021

Abstract

Disclosed herein are methods and compositions for modifying TCR genes, using nucleases (zinc finger nucleases or TAL nucleases) to modify TCR genes.

First claim

What is claimed is:

1. A population of isolated T-lymphocytes comprising:
(i) CD62L+stem memory (TSCM) cells, central memory T cells (TCM), T effector memory cell (TEM) and terminal effector T cells (TEMRA);
(ii) endogenous T cell receptor (TCR) alpha (TRAC) or TCR beta (TRBC) genes;
(iii) one or more mRNAs encoding a pair of zinc finger nucleases (ZFNs) specific for the TRAC gene, each zinc finger nuclease comprising a zinc finger protein that binds to a target sequence in an exon of the TRAC gene and an engineered cleavage half-domain, wherein the zinc protein comprises 4, 5, or 6 zinc finger domains ordered F1 to F4, F1 to F5 or F1 to F6, the zinc finger domains comprising the recognition helix regions in order shown in a single row of the following Table:

F1
F2
F3
F4
F5
F6

QSGDLTR
QRTHLKA
QSGDRNK
DRSNLSR
RSDALTQ
N/A

(SEQ ID 
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 2) 
NO: 3)
NO: 4)
NO: 5)
NO: 6)

TSGSLSR
QSSVRNS
RSDNLST
DRSALAR
LKQNLDA
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 8) 
NO: 9)
NO: 10)
NO: 11)
NO: 12)

DRSALSR
QSGHLSR
DRSDLSR
RSDALSR
DRSDLSR
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 14)
NO: 15)
NO: 16)
NO: 17)
NO: 16)

DRSNLSR
QKTSLQA
DRSALSR
QSGNLAR
GKEELNE
RSSDLSR

(SEQ ID
(SEQ ID 
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 5)
NO: 19)
NO: 14)
NO: 20)
NO: 21)
NO: 22)

GNVDLIE
RSSNLSR
RSDALSV
DSSHRTR
WRSCRSA
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 24)
NO: 25)
NO: 26) 
NO: 27)
NO: 28)

DSSDRKK
RSDNLSV
RRFILRG
QSGDLTR
TSGSLTR
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 30)
NO: 31)
NO: 32)
NO: 2)
NO: 33)

QSGDLTR
QTSTLSK
QSGHLSR
DRSDLSR
RSDALAR
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 2)
NO: 35)
NO: 15)
NO: 16)
NO: 36)

QSGDLTR
WRSSLAS
QSGDLTR
HKWVLRQ
DRSNLTR
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 2)
NO: 38)
NO: 2)
NO: 39)
NO: 40)

QSGDLTR
QWGTRYR
ERGTLAR
RSDNLRE
QSGDLTR
TSGSLTR

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 2)
NO: 42)
NO: 43)
NO: 44)
NO: 2)
NO: 33)

RSAHLSR
DRSDLSR
RSDHLSV
QNNHRIT
N/A
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 46)
NO: 16)
NO: 47)
NO: 48)

QRSNLVR
RNDDRKK
TSGNLTR
TSANLSR
N/A
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 50)
NO: 51)
NO: 52)
NO: 53)

DRSTLRQ
QRSNLVR
RNDDRKK
RSAHLSR
QSGHLSR
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO: 55)
NO: 50)
NO: 51)
NO: 46)
NO: 15)

QRSNLVR
RNDDRKK
QSGHLAR
QSGHLSR
N/A
N/A

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

Assignees

• Sangamo Therapeutics Inc
• Ospedale San Raffaele Srl

Inventors

• Gregory Philip D
• Holmes Michael C
• Paschon David
• Zhang Lei
• Bonini Maria Chiara
• Genovese Pietro
• Magnani Zulma
• Mastaglio Sara
• Naldini Luigi

Classifications

• A61K40/4269

  NY-ESO · CPC title

• A61K40/4243

  Wilms tumor 1 [WT1] · CPC title

• A61K40/32

  T-cell receptors [TCR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/48

  Blood cells, e.g. leukemia or lymphoma · CPC title