               SEQUENCE LISTING

<110> Medizinische Hochschule Hannover

<120> HLA-DR-specific gamma delta TCR construct and use thereof

<130> 13210

<150> EP21163630
<151> 2021-03-19

<160> 38

<170> BiSSAP 1.3.6

<210> 1
<211> 23
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD CDR3 (TCR04)

<400> 1
Ala Leu Gly Glu Leu Tyr Val Pro Leu Tyr Trp Gly Pro Thr Pro Arg 
1               5                   10                  15      
Thr Thr Asp Lys Leu Ile Phe 
            20              

<210> 2
<211> 23
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD CDR3 (consensus sequence)

<220> 
<221> VARIANT
<222> 7
<223> X can be any amino acid, preferably, the amino acid in the
      variable position X is a hydrophobic amino acid, e.g., a
      non-aromatic hydrophobic amino acid (i.e., Val, Ala, Gly, Ile,
      Leu, Val or Met)

<400> 2
Ala Leu Gly Glu Leu Tyr Xaa Pro Leu Tyr Trp Gly Pro Thr Pro Arg 
1               5                   10                  15      
Thr Thr Asp Lys Leu Ile Phe 
            20              

<210> 3
<211> 23
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD CDR3 (TCR05)

<400> 3
Ala Leu Gly Glu Leu Tyr Gly Pro Leu Tyr Trp Gly Pro Thr Pro Arg 
1               5                   10                  15      
Thr Thr Asp Lys Leu Ile Phe 
            20              

<210> 4
<211> 23
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD CDR3 (with ALA)

<400> 4
Ala Leu Gly Glu Leu Tyr Ala Pro Leu Tyr Trp Gly Pro Thr Pro Arg 
1               5                   10                  15      
Thr Thr Asp Lys Leu Ile Phe 
            20              

<210> 5
<211> 7
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD CDR1

<400> 5
Thr Ser Trp Trp Ser Tyr Tyr 
1               5           

<210> 6
<400> 6
000

<210> 7
<211> 94
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD variable region (TCR04)

<400> 7
Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser Met Pro Val Arg 
1               5                   10                  15      
Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser Trp Trp Ser Tyr 
            20                  25                  30          
Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu Met Ile Phe Leu 
        35                  40                  45              
Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser Gly Arg Tyr Ser 
    50                  55                  60                  
Val Asn Phe Lys Lys Ala Ala Lys Ser Val Ala Leu Thr Ile Ser Ala 
65                  70                  75                  80  
Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala Leu Gly 
                85                  90                  

<210> 8
<211> 282
<212> DNA
<213> Homo sapiens


<220> 
<223> TRD variable region na (TCR04)

<400> 8
gcccagaagg tgacccaggc ccagtcctcc gtgtccatgc ctgtgagaaa ggccgtgacc     60

ctgaattgtc tgtacgagac aagctggtgg tcctactaca tcttttggta caagcagctg    120

ccttccaagg agatgatctt tctgatcaga cagggcagcg atgagcagaa cgccaagtcc    180

ggcaggtact ccgtgaactt taagaaggcc gccaagtccg tggccctgac aatcagcgcc    240

ctgcagctgg aggatagcgc caagtacttt tgcgccctgg gc                       282


<210> 9
<211> 10
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG CDR3

<400> 9
Ala Thr Trp Asp Arg Gly Arg Lys Leu Phe 
1               5                   10  

<210> 10
<211> 10
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG CDR3

<400> 10
Ala Thr Trp Asp Arg Gly Arg Lys Leu Phe 
1               5                   10  

<210> 11
<211> 6
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG CDR1

<400> 11
Val Thr Asn Thr Phe Tyr 
1               5       

<210> 12
<211> 8
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG CDR2

<400> 12
Tyr Asp Val Ser Thr Ala Arg Asp 
1               5               

<210> 13
<211> 100
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG variable region

<400> 13
Ser Ser Asn Leu Glu Gly Arg Thr Lys Ser Val Thr Arg Gln Thr Gly 
1               5                   10                  15      
Ser Ser Ala Glu Ile Thr Cys Asp Leu Thr Val Thr Asn Thr Phe Tyr 
            20                  25                  30          
Ile His Trp Tyr Leu His Gln Glu Gly Lys Ala Pro Gln Arg Leu Leu 
        35                  40                  45              
Tyr Tyr Asp Val Ser Thr Ala Arg Asp Val Leu Glu Ser Gly Leu Ser 
    50                  55                  60                  
Pro Gly Lys Tyr Tyr Thr His Thr Pro Arg Arg Trp Ser Trp Ile Leu 
65                  70                  75                  80  
Arg Leu Gln Asn Leu Ile Glu Asn Asp Ser Gly Val Tyr Tyr Cys Ala 
                85                  90                  95      
Thr Trp Asp Arg 
            100 

<210> 14
<211> 300
<212> DNA
<213> Homo sapiens


<220> 
<223> TRG variable region nucleic acid

<400> 14
tccagcaatc tggagggcag gacaaagtcc gtgacaaggc agacaggctc cagcgccgag     60

atcacatgcg acctgacagt gacaaatacc ttctacatcc actggtacct gcaccaggag    120

ggcaaggccc ctcagagact gctgtactac gacgtgagca cagccaggga cgtgctggag    180

tccggcctgt cccctggcaa gtactacaca cacaccccta gaagatggag ctggatcttg    240

agactgcaga atctgatcga gaacgattcc ggcgtgtact actgcgccac atgggataga    300


<210> 15
<211> 305
<212> PRT
<213> Homo sapiens


<220> 
<223> TRG full length (TCR04, aa)

<400> 15
Met Gly Trp Ala Leu Val Val Leu Leu Ala Phe Leu Ser Pro Ala Ser 
1               5                   10                  15      
Gln Lys Ser Ser Asn Leu Glu Gly Arg Thr Lys Ser Val Thr Arg Gln 
            20                  25                  30          
Thr Gly Ser Ser Ala Glu Ile Thr Cys Asp Leu Thr Val Thr Asn Thr 
        35                  40                  45              
Phe Tyr Ile His Trp Tyr Leu His Gln Glu Gly Lys Ala Pro Gln Arg 
    50                  55                  60                  
Leu Leu Tyr Tyr Asp Val Ser Thr Ala Arg Asp Val Leu Glu Ser Gly 
65                  70                  75                  80  
Leu Ser Pro Gly Lys Tyr Tyr Thr His Thr Pro Arg Arg Trp Ser Trp 
                85                  90                  95      
Ile Leu Arg Leu Gln Asn Leu Ile Glu Asn Asp Ser Gly Val Tyr Tyr 
            100                 105                 110         
Cys Ala Thr Trp Asp Arg Gly Arg Lys Leu Phe Gly Ser Gly Thr Thr 
        115                 120                 125             
Leu Val Val Thr Asp Lys Gln Leu Asp Ala Asp Val Ser Pro Lys Pro 
    130                 135                 140                 
Thr Ile Phe Leu Pro Ser Ile Ala Glu Thr Lys Leu Gln Lys Ala Gly 
145                 150                 155                 160 
Thr Tyr Leu Cys Leu Leu Glu Lys Phe Phe Pro Asp Val Ile Lys Ile 
                165                 170                 175     
His Trp Gln Glu Lys Lys Ser Asn Thr Ile Leu Gly Ser Gln Glu Gly 
            180                 185                 190         
Asn Thr Met Lys Thr Asn Asp Thr Tyr Met Lys Phe Ser Trp Leu Thr 
        195                 200                 205             
Val Pro Glu Lys Ser Leu Asp Lys Glu His Arg Cys Ile Val Arg His 
    210                 215                 220                 
Glu Asn Asn Lys Asn Gly Val Asp Gln Glu Ile Ile Phe Pro Pro Ile 
225                 230                 235                 240 
Lys Thr Asp Val Ile Thr Met Asp Pro Lys Asp Asn Cys Ser Lys Asp 
                245                 250                 255     
Ala Asn Asp Thr Leu Leu Leu Gln Leu Thr Asn Thr Ser Ala Tyr Tyr 
            260                 265                 270         
Met Tyr Leu Leu Leu Leu Leu Lys Ser Val Val Tyr Phe Ala Ile Ile 
        275                 280                 285             
Thr Cys Cys Leu Leu Arg Arg Thr Ala Phe Cys Cys Asn Gly Glu Lys 
    290                 295                 300                 
Ser 
305 

<210> 16
<211> 298
<212> PRT
<213> Homo sapiens


<220> 
<223> TRD full length (aa)

<400> 16
Met Val Phe Ser Ser Leu Leu Cys Val Phe Val Ala Phe Ser Tyr Ser 
1               5                   10                  15      
Gly Ser Ser Val Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser 
            20                  25                  30          
Met Pro Val Arg Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser 
        35                  40                  45              
Trp Trp Ser Tyr Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu 
    50                  55                  60                  
Met Ile Phe Leu Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser 
65                  70                  75                  80  
Gly Arg Tyr Ser Val Asn Phe Lys Lys Ala Ala Lys Ser Val Ala Leu 
                85                  90                  95      
Thr Ile Ser Ala Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala 
            100                 105                 110         
Leu Gly Glu Leu Tyr Val Pro Leu Tyr Trp Gly Pro Thr Pro Arg Thr 
        115                 120                 125             
Thr Asp Lys Leu Ile Phe Gly Lys Gly Thr Arg Val Thr Val Glu Pro 
    130                 135                 140                 
Arg Ser Gln Pro His Thr Lys Pro Ser Val Phe Val Met Lys Asn Gly 
145                 150                 155                 160 
Thr Asn Val Ala Cys Leu Val Lys Glu Phe Tyr Pro Lys Asp Ile Arg 
                165                 170                 175     
Ile Asn Leu Val Ser Ser Lys Lys Ile Thr Glu Phe Asp Pro Ala Ile 
            180                 185                 190         
Val Ile Ser Pro Ser Gly Lys Tyr Asn Ala Val Lys Leu Gly Lys Tyr 
        195                 200                 205             
Glu Asp Ser Asn Ser Val Thr Cys Ser Val Gln His Asp Asn Lys Thr 
    210                 215                 220                 
Val His Ser Thr Asp Phe Glu Val Lys Thr Asp Ser Thr Asp His Val 
225                 230                 235                 240 
Lys Pro Lys Glu Thr Glu Asn Thr Lys Gln Pro Ser Lys Ser Cys His 
                245                 250                 255     
Lys Pro Lys Ala Ile Val His Thr Glu Lys Val Asn Met Met Ser Leu 
            260                 265                 270         
Thr Val Leu Gly Leu Arg Met Leu Phe Ala Lys Thr Val Ala Val Asn 
        275                 280                 285             
Phe Leu Leu Thr Ala Lys Leu Phe Phe Leu 
    290                 295             

<210> 17
<211> 303
<212> PRT
<213> Homo sapiens


<220> 
<221> SIGNAL
<222> 1..20
<223> signal peptide for expression in insect cells

<220> 
<223> TRD soluble (TCR04, aa)

<220> 
<221> BINDING
<222> 267..303
<223> Twin-Strep-Tag

<400> 17
Met Lys Leu Cys Ile Leu Leu Ala Val Val Ala Phe Val Gly Leu Ser 
1               5                   10                  15      
Leu Gly Arg Ser Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser 
            20                  25                  30          
Met Pro Val Arg Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser 
        35                  40                  45              
Trp Trp Ser Tyr Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu 
    50                  55                  60                  
Met Ile Phe Leu Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser 
65                  70                  75                  80  
Gly Arg Tyr Ser Val Asn Phe Lys Lys Ala Ala Lys Ser Val Ala Leu 
                85                  90                  95      
Thr Ile Ser Ala Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala 
            100                 105                 110         
Leu Gly Glu Leu Tyr Val Pro Leu Tyr Trp Gly Pro Thr Pro Arg Thr 
        115                 120                 125             
Thr Asp Lys Leu Ile Phe Gly Lys Gly Thr Arg Val Thr Val Glu Pro 
    130                 135                 140                 
Arg Ser Gln Pro His Thr Lys Pro Ser Val Phe Val Met Lys Asn Gly 
145                 150                 155                 160 
Thr Asn Val Ala Cys Leu Val Lys Glu Phe Tyr Pro Lys Asp Ile Arg 
                165                 170                 175     
Ile Asn Leu Val Ser Ser Lys Lys Ile Thr Glu Phe Asp Pro Ala Ile 
            180                 185                 190         
Val Ile Ser Pro Ser Gly Lys Tyr Asn Ala Val Lys Leu Gly Lys Tyr 
        195                 200                 205             
Glu Asp Ser Asn Ser Val Thr Cys Ser Val Gln His Asp Asn Lys Thr 
    210                 215                 220                 
Val His Ser Thr Asp Phe Glu Val Lys Thr Asp Ser Thr Asp His Val 
225                 230                 235                 240 
Lys Pro Lys Glu Thr Glu Asn Thr Lys Gln Pro Ser Lys Ser Cys His 
                245                 250                 255     
Lys Pro Lys Ala Ile Val His Thr Glu Lys Ser Arg Asp Asp Asp Asp 
            260                 265                 270         
Lys Ala Gly Trp Ser His Pro Gln Phe Glu Lys Gly Gly Gly Ser Gly 
        275                 280                 285             
Gly Gly Ser Gly Gly Gly Ser Trp Ser His Pro Gln Phe Glu Lys 
    290                 295                 300             

<210> 18
<211> 303
<212> PRT
<213> Homo sapiens


<220> 
<221> SIGNAL
<222> 1..20
<223> signal peptide for expression in insect cells

<220> 
<223> TRG soluble (aa)

<220> 
<221> BINDING
<222> 267..303
<223> Twin-Strep-Tag

<400> 18
Met Lys Leu Cys Ile Leu Leu Ala Val Val Ala Phe Val Gly Leu Ser 
1               5                   10                  15      
Leu Gly Arg Ser Ser Ser Asn Leu Glu Gly Arg Thr Lys Ser Val Thr 
            20                  25                  30          
Arg Gln Thr Gly Ser Ser Ala Glu Ile Thr Cys Asp Leu Thr Val Thr 
        35                  40                  45              
Asn Thr Phe Tyr Ile His Trp Tyr Leu His Gln Glu Gly Lys Ala Pro 
    50                  55                  60                  
Gln Arg Leu Leu Tyr Tyr Asp Val Ser Thr Ala Arg Asp Val Leu Glu 
65                  70                  75                  80  
Ser Gly Leu Ser Pro Gly Lys Tyr Tyr Thr His Thr Pro Arg Arg Trp 
                85                  90                  95      
Ser Trp Ile Leu Arg Leu Gln Asn Leu Ile Glu Asn Asp Ser Gly Val 
            100                 105                 110         
Tyr Tyr Cys Ala Thr Trp Asp Arg Gly Arg Lys Leu Phe Gly Ser Gly 
        115                 120                 125             
Thr Thr Leu Val Val Thr Asp Lys Gln Leu Asp Ala Asp Val Ser Pro 
    130                 135                 140                 
Lys Pro Thr Ile Phe Leu Pro Ser Ile Ala Glu Thr Lys Leu Gln Lys 
145                 150                 155                 160 
Ala Gly Thr Tyr Leu Cys Leu Leu Glu Lys Phe Phe Pro Asp Val Ile 
                165                 170                 175     
Lys Ile His Trp Gln Glu Lys Lys Ser Asn Thr Ile Leu Gly Ser Gln 
            180                 185                 190         
Glu Gly Asn Thr Met Lys Thr Asn Asp Thr Tyr Met Lys Phe Ser Trp 
        195                 200                 205             
Leu Thr Val Pro Glu Lys Ser Leu Asp Lys Glu His Arg Cys Ile Val 
    210                 215                 220                 
Arg His Glu Asn Asn Lys Asn Gly Val Asp Gln Glu Ile Ile Phe Pro 
225                 230                 235                 240 
Pro Ile Lys Thr Asp Val Ile Thr Met Asp Pro Lys Asp Asn Cys Ser 
                245                 250                 255     
Lys Asp Ala Asn Asp Thr Leu Leu Leu Gln Ser Arg Asp Asp Asp Asp 
            260                 265                 270         
Lys Ala Gly Trp Ser His Pro Gln Phe Glu Lys Gly Gly Gly Ser Gly 
        275                 280                 285             
Gly Gly Ser Gly Gly Gly Ser Trp Ser His Pro Gln Phe Glu Lys 
    290                 295                 300             

<210> 19
<211> 25
<212> DNA
<213> Artificial Sequence


<220> 
<223> Vgamma3-forward primer

<400> 19
gaagatcttc cagcaatctg gaggg                                          25


<210> 20
<211> 26
<212> DNA
<213> Artificial Sequence


<220> 
<223> Vgamma3-reverse primer

<400> 20
gctctagact gcagcagcag ggtatc                                         26


<210> 21
<211> 27
<212> DNA
<213> Artificial Sequence


<220> 
<223> Vdelta1-forward primer

<400> 21
gaagatctgc ccagaaggtg acccagg                                        27


<210> 22
<211> 29
<212> DNA
<213> Artificial Sequence


<220> 
<223> Vdelta1-reverse primer

<400> 22
gctctagact tctctgtgtg cacgatggc                                      29


<210> 23
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> TRB KO in JE6.1 gRNA1

<400> 23
ggctcaaaca cagcgacctc                                                20


<210> 24
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> TRB KO in JE6.1 gRNA2

<400> 24
ggctctcgga gaatgacgag                                                20


<210> 25
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> HLA-DRA KO in Raji

<400> 25
tgcattggcc aacatagctg                                                20


<210> 26
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> RFXAP KO in Raji

<400> 26
acaccgcaac aagatgtaca                                                20


<210> 27
<211> 16
<212> PRT
<213> Homo sapiens


<220> 
<223> CDR3 of TCR02

<400> 27
Ala Leu Gly Glu Leu Pro Arg His His Leu Thr Asp Lys Leu Ile Phe 
1               5                   10                  15      



<210> 28
<211> 15
<212> PRT
<213> Homo sapiens


<220> 
<223> peptide loaded on HLA-DR - CLIP87-101

<400> 28
Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala 
1               5                   10                  15  

<210> 29
<211> 14
<212> PRT
<213> Cytomegalovirus


<220> 
<223> peptide loaded on HLA-DR - CMV pp65 116-129

<400> 29
Leu Pro Leu Lys Met Leu Asn Ile Pro Ser Ile Asn Val His 
1               5                   10                  

<210> 30
<211> 16
<212> PRT
<213> Homo sapiens


<220> 
<223> peptide loaded on HLA-DR - MAGE-A3 267-282

<400> 30
Ala Cys Tyr Glu Phe Leu Trp Gly Pro Arg Ala Leu Val Glu Thr Ser 
1               5                   10                  15      



<210> 31
<211> 13
<212> PRT
<213> Human herpesvirus 4


<220> 
<223> peptide loaded on HLA-DR - EBV EBNA1 515-527

<400> 31
Thr Ser Leu Tyr Asn Leu Arg Arg Gly Thr Ala Leu Ala 
1               5                   10              

<210> 32
<211> 11
<212> PRT
<213> Human herpesvirus 4


<220> 
<223> peptide loaded on HLA-DR - EBV EBNA2 280-290

<400> 32
Thr Val Phe Tyr Asn Ile Pro Pro Met Pro Leu 
1               5                   10      

<210> 33
<211> 15
<212> PRT
<213> Dermatophagoides pteronyssinus


<220> 
<223> peptide loaded on HLA-DR - house dust mit DR p1 96-110

<400> 33
Leu Arg Gln Met Arg Thr Val Thr Pro Ile Arg Met Gln Gly Gly 
1               5                   10                  15  

<210> 34
<211> 15
<212> PRT
<213> Cytomegalovirus


<220> 
<223> peptide loaded on HLA-DR - HCMV IE1

<400> 34
Thr Arg Arg Gly Arg Val Lys Ile Asp Glu Val Ser Arg Met Phe 
1               5                   10                  15  

<210> 35
<211> 15
<212> PRT
<213> Cytomegalovirus


<220> 
<223> peptide loaded on HLA-DR - HCMV IE2

<400> 35
Val Lys Gln Ile Lys Val Arg Val Asp Met Val Arg His Arg Ile 
1               5                   10                  15  

<210> 36
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> sgRNA for HLA-DRB KO

<400> 36
gagtactgga acagccaga                                                 19


<210> 37
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> crRNA for knock-in of HLA-DRB gene

<400> 37
ggcggcctga tgccgagtac                                                20


<210> 38
<211> 122
<212> DNA
<213> Artificial Sequence


<220> 
<223> ssDNA oligomer for knock-in of HLA-DRB gene

<400> 38
ttcgacagcg acgtggggga gttccgggcg gtgacggagc tgggccgccc agacgccgaa     60

tattggaaca gccagaagga cctcctggag cagaagcggg gccgggtgga caactactgc    120

aa                                                                   122


