                         SEQUENCE LISTING

<110>  Yeda Research and Development Co. Ltd.
       WALLACH, David
       YOON, Seongmin
 
<120>  CONTROLLING UBIQUITINATION OF MLKL FOR TREATMENT OF DISEASE

<130>  90063

<150>  IL 279559
<151>  2020-12-17

<160>  14    

<170>  PatentIn version 3.5

<210>  1
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  gRNA sequences

<400>  1
gttcctatgc ctcgggcgcg t                                                 21


<210>  2
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  gRNA sequences

<400>  2
acgcgcccga ggcataggaa c                                                 21


<210>  3
<211>  28
<212>  PRT
<213>  Artificial sequence

<220>
<223>   double strep tag

<400>  3

Trp Ser His Pro Gln Phe Glu Lys Gly Gly Gly Ser Gly Gly Gly Ser 
1               5                   10                  15      


Gly Gly Gly Ser Trp Ser His Pro Gln Phe Glu Lys 
            20                  25              


<210>  4
<211>  10
<212>  PRT
<213>  Artificial sequence

<220>
<223>  myc tag

<400>  4

Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 
1               5                   10  


<210>  5
<211>  8
<212>  PRT
<213>  Artificial sequence

<220>
<223>  FLAG-tag

<400>  5

Asp Tyr Lys Asp Asp Asp Asp Lys 
1               5               


<210>  6
<211>  471
<212>  PRT
<213>  homo sapiens

<400>  6

Met Glu Asn Leu Lys His Ile Ile Thr Leu Gly Gln Val Ile His Lys 
1               5                   10                  15      


Arg Cys Glu Glu Met Lys Tyr Cys Lys Lys Gln Cys Arg Arg Leu Gly 
            20                  25                  30          


His Arg Val Leu Gly Leu Ile Lys Pro Leu Glu Met Leu Gln Asp Gln 
        35                  40                  45              


Gly Lys Arg Ser Val Pro Ser Glu Lys Leu Thr Thr Ala Met Asn Arg 
    50                  55                  60                  


Phe Lys Ala Ala Leu Glu Glu Ala Asn Gly Glu Ile Glu Lys Phe Ser 
65                  70                  75                  80  


Asn Arg Ser Asn Ile Cys Arg Phe Leu Thr Ala Ser Gln Asp Lys Ile 
                85                  90                  95      


Leu Phe Lys Asp Val Asn Arg Lys Leu Ser Asp Val Trp Lys Glu Leu 
            100                 105                 110         


Ser Leu Leu Leu Gln Val Glu Gln Arg Met Pro Val Ser Pro Ile Ser 
        115                 120                 125             


Gln Gly Ala Ser Trp Ala Gln Glu Asp Gln Gln Asp Ala Asp Glu Asp 
    130                 135                 140                 


Arg Arg Ala Phe Gln Met Leu Arg Arg Asp Asn Glu Lys Ile Glu Ala 
145                 150                 155                 160 


Ser Leu Arg Arg Leu Glu Ile Asn Met Lys Glu Ile Lys Glu Thr Leu 
                165                 170                 175     


Arg Gln Tyr Leu Pro Pro Lys Cys Met Gln Glu Ile Pro Gln Glu Gln 
            180                 185                 190         


Ile Lys Glu Ile Lys Lys Glu Gln Leu Ser Gly Ser Pro Trp Ile Leu 
        195                 200                 205             


Leu Arg Glu Asn Glu Val Ser Thr Leu Tyr Lys Gly Glu Tyr His Arg 
    210                 215                 220                 


Ala Pro Val Ala Ile Lys Val Phe Lys Lys Leu Gln Ala Gly Ser Ile 
225                 230                 235                 240 


Ala Ile Val Arg Gln Thr Phe Asn Lys Glu Ile Lys Thr Met Lys Lys 
                245                 250                 255     


Phe Glu Ser Pro Asn Ile Leu Arg Ile Phe Gly Ile Cys Ile Asp Glu 
            260                 265                 270         


Thr Val Thr Pro Pro Gln Phe Ser Ile Val Met Glu Tyr Cys Glu Leu 
        275                 280                 285             


Gly Thr Leu Arg Glu Leu Leu Asp Arg Glu Lys Asp Leu Thr Leu Gly 
    290                 295                 300                 


Lys Arg Met Val Leu Val Leu Gly Ala Ala Arg Gly Leu Tyr Arg Leu 
305                 310                 315                 320 


His His Ser Glu Ala Pro Glu Leu His Gly Lys Ile Arg Ser Ser Asn 
                325                 330                 335     


Phe Leu Val Thr Gln Gly Tyr Gln Val Lys Leu Ala Gly Phe Glu Leu 
            340                 345                 350         


Arg Lys Thr Gln Thr Ser Met Ser Leu Gly Thr Thr Arg Glu Lys Thr 
        355                 360                 365             


Asp Arg Val Lys Ser Thr Ala Tyr Leu Ser Pro Gln Glu Leu Glu Asp 
    370                 375                 380                 


Val Phe Tyr Gln Tyr Asp Val Lys Ser Glu Ile Tyr Ser Phe Gly Ile 
385                 390                 395                 400 


Val Leu Trp Glu Ile Ala Thr Gly Asp Ile Pro Phe Gln Gly Cys Asn 
                405                 410                 415     


Ser Glu Lys Ile Arg Lys Leu Val Ala Val Lys Arg Gln Gln Glu Pro 
            420                 425                 430         


Leu Gly Glu Asp Cys Pro Ser Glu Leu Arg Glu Ile Ile Asp Glu Cys 
        435                 440                 445             


Arg Ala His Asp Pro Ser Val Arg Pro Ser Val Asp Glu Ile Leu Lys 
    450                 455                 460                 


Lys Leu Ser Thr Phe Ser Lys 
465                 470     


<210>  7
<211>  862
<212>  PRT
<213>  homo sapiens

<400>  7

Met Ser Asp Ser Gly Ser Gln Leu Gly Ser Met Gly Ser Leu Thr Met 
1               5                   10                  15      


Lys Ser Gln Leu Gln Ile Thr Val Ile Ser Ala Lys Leu Lys Glu Asn 
            20                  25                  30          


Lys Lys Asn Trp Phe Gly Pro Ser Pro Tyr Val Glu Val Thr Val Asp 
        35                  40                  45              


Gly Gln Ser Lys Lys Thr Glu Lys Cys Asn Asn Thr Asn Ser Pro Lys 
    50                  55                  60                  


Trp Lys Gln Pro Leu Thr Val Ile Val Thr Pro Val Ser Lys Leu His 
65                  70                  75                  80  


Phe Arg Val Trp Ser His Gln Thr Leu Lys Ser Asp Val Leu Leu Gly 
                85                  90                  95      


Thr Ala Ala Leu Asp Ile Tyr Glu Thr Leu Lys Ser Asn Asn Met Lys 
            100                 105                 110         


Leu Glu Glu Val Val Val Thr Leu Gln Leu Gly Gly Asp Lys Glu Pro 
        115                 120                 125             


Thr Glu Thr Ile Gly Asp Leu Ser Ile Cys Leu Asp Gly Leu Gln Leu 
    130                 135                 140                 


Glu Ser Glu Val Val Thr Asn Gly Glu Thr Thr Cys Ser Glu Ser Ala 
145                 150                 155                 160 


Ser Gln Asn Asp Asp Gly Ser Arg Ser Lys Asp Glu Thr Arg Val Ser 
                165                 170                 175     


Thr Asn Gly Ser Asp Asp Pro Glu Asp Ala Gly Ala Gly Glu Asn Arg 
            180                 185                 190         


Arg Val Ser Gly Asn Asn Ser Pro Ser Leu Ser Asn Gly Gly Phe Lys 
        195                 200                 205             


Pro Ser Arg Pro Pro Arg Pro Ser Arg Pro Pro Pro Pro Thr Pro Arg 
    210                 215                 220                 


Arg Pro Ala Ser Val Asn Gly Ser Pro Ser Ala Thr Ser Glu Ser Asp 
225                 230                 235                 240 


Gly Ser Ser Thr Gly Ser Leu Pro Pro Thr Asn Thr Asn Thr Asn Thr 
                245                 250                 255     


Ser Glu Gly Ala Thr Ser Gly Leu Ile Ile Pro Leu Thr Ile Ser Gly 
            260                 265                 270         


Gly Ser Gly Pro Arg Pro Leu Asn Pro Val Thr Gln Ala Pro Leu Pro 
        275                 280                 285             


Pro Gly Trp Glu Gln Arg Val Asp Gln His Gly Arg Val Tyr Tyr Val 
    290                 295                 300                 


Asp His Val Glu Lys Arg Thr Thr Trp Asp Arg Pro Glu Pro Leu Pro 
305                 310                 315                 320 


Pro Gly Trp Glu Arg Arg Val Asp Asn Met Gly Arg Ile Tyr Tyr Val 
                325                 330                 335     


Asp His Phe Thr Arg Thr Thr Thr Trp Gln Arg Pro Thr Leu Glu Ser 
            340                 345                 350         


Val Arg Asn Tyr Glu Gln Trp Gln Leu Gln Arg Ser Gln Leu Gln Gly 
        355                 360                 365             


Ala Met Gln Gln Phe Asn Gln Arg Phe Ile Tyr Gly Asn Gln Asp Leu 
    370                 375                 380                 


Phe Ala Thr Ser Gln Ser Lys Glu Phe Asp Pro Leu Gly Pro Leu Pro 
385                 390                 395                 400 


Pro Gly Trp Glu Lys Arg Thr Asp Ser Asn Gly Arg Val Tyr Phe Val 
                405                 410                 415     


Asn His Asn Thr Arg Ile Thr Gln Trp Glu Asp Pro Arg Ser Gln Gly 
            420                 425                 430         


Gln Leu Asn Glu Lys Pro Leu Pro Glu Gly Trp Glu Met Arg Phe Thr 
        435                 440                 445             


Val Asp Gly Ile Pro Tyr Phe Val Asp His Asn Arg Arg Thr Thr Thr 
    450                 455                 460                 


Tyr Ile Asp Pro Arg Thr Gly Lys Ser Ala Leu Asp Asn Gly Pro Gln 
465                 470                 475                 480 


Ile Ala Tyr Val Arg Asp Phe Lys Ala Lys Val Gln Tyr Phe Arg Phe 
                485                 490                 495     


Trp Cys Gln Gln Leu Ala Met Pro Gln His Ile Lys Ile Thr Val Thr 
            500                 505                 510         


Arg Lys Thr Leu Phe Glu Asp Ser Phe Gln Gln Ile Met Ser Phe Ser 
        515                 520                 525             


Pro Gln Asp Leu Arg Arg Arg Leu Trp Val Ile Phe Pro Gly Glu Glu 
    530                 535                 540                 


Gly Leu Asp Tyr Gly Gly Val Ala Arg Glu Trp Phe Phe Leu Leu Ser 
545                 550                 555                 560 


His Glu Val Leu Asn Pro Met Tyr Cys Leu Phe Glu Tyr Ala Gly Lys 
                565                 570                 575     


Asp Asn Tyr Cys Leu Gln Ile Asn Pro Ala Ser Tyr Ile Asn Pro Asp 
            580                 585                 590         


His Leu Lys Tyr Phe Arg Phe Ile Gly Arg Phe Ile Ala Met Ala Leu 
        595                 600                 605             


Phe His Gly Lys Phe Ile Asp Thr Gly Phe Ser Leu Pro Phe Tyr Lys 
    610                 615                 620                 


Arg Ile Leu Asn Lys Pro Val Gly Leu Lys Asp Leu Glu Ser Ile Asp 
625                 630                 635                 640 


Pro Glu Phe Tyr Asn Ser Leu Ile Trp Val Lys Glu Asn Asn Ile Glu 
                645                 650                 655     


Glu Cys Asp Leu Glu Met Tyr Phe Ser Val Asp Lys Glu Ile Leu Gly 
            660                 665                 670         


Glu Ile Lys Ser His Asp Leu Lys Pro Asn Gly Gly Asn Ile Leu Val 
        675                 680                 685             


Thr Glu Glu Asn Lys Glu Glu Tyr Ile Arg Met Val Ala Glu Trp Arg 
    690                 695                 700                 


Leu Ser Arg Gly Val Glu Glu Gln Thr Gln Ala Phe Phe Glu Gly Phe 
705                 710                 715                 720 


Asn Glu Ile Leu Pro Gln Gln Tyr Leu Gln Tyr Phe Asp Ala Lys Glu 
                725                 730                 735     


Leu Glu Val Leu Leu Cys Gly Met Gln Glu Ile Asp Leu Asn Asp Trp 
            740                 745                 750         


Gln Arg His Ala Ile Tyr Arg His Tyr Ala Arg Thr Ser Lys Gln Ile 
        755                 760                 765             


Met Trp Phe Trp Gln Phe Val Lys Glu Ile Asp Asn Glu Lys Arg Met 
    770                 775                 780                 


Arg Leu Leu Gln Phe Val Thr Gly Thr Cys Arg Leu Pro Val Gly Gly 
785                 790                 795                 800 


Phe Ala Asp Leu Met Gly Ser Asn Gly Pro Gln Lys Phe Cys Ile Glu 
                805                 810                 815     


Lys Val Gly Lys Glu Asn Trp Leu Pro Arg Ser His Thr Cys Phe Asn 
            820                 825                 830         


Arg Leu Asp Leu Pro Pro Tyr Lys Ser Tyr Glu Gln Leu Lys Glu Lys 
        835                 840                 845             


Leu Leu Phe Ala Ile Glu Glu Thr Glu Gly Phe Gly Gln Glu 
    850                 855                 860         


<210>  8
<211>  14
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS

<400>  8

Phe Lys Ala Ala Leu Glu Glu Ala Asn Gly Glu Ile Glu Lys 
1               5                   10                  


<210>  9
<211>  14
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS


<220>
<221>  MOD_RES
<222>  (12)..(12)
<223>  may be ubiquitinated

<400>  9

Leu His His Ser Glu Ala Pro Glu Leu His Gly Lys Ile Arg 
1               5                   10                  


<210>  10
<211>  17
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS


<220>
<221>  MOD_RES
<222>  (10)..(10)
<223>  may be oxidized

<220>
<221>  MOD_RES
<222>  (16)..(16)
<223>  may be ubiquitinated

<400>  10

Val Leu Gly Leu Ile Lys Pro Leu Glu Met Leu Gln Asp Gln Gly Lys 
1               5                   10                  15      


Arg 
    


<210>  11
<211>  12
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS

<400>  11

Ala Ala Leu Glu Glu Ala Asn Gly Glu Ile Glu Lys 
1               5                   10          


<210>  12
<211>  12
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS


<220>
<221>  MOD_RES
<222>  (12)..(12)
<223>  may be ubiquitinated

<400>  12

Leu His His Ser Glu Ala Pro Glu Leu His Gly Lys 
1               5                   10          


<210>  13
<211>  16
<212>  PRT
<213>  Artificial sequence

<220>
<223>  isolated peptides analyzed by MS/MS


<220>
<221>  MOD_RES
<222>  (10)..(10)
<223>  oxidized

<220>
<221>  MOD_RES
<222>  (16)..(16)
<223>  may be ubiquitinated

<400>  13

Val Leu Gly Leu Ile Lys Pro Leu Glu Met Leu Gln Asp Gln Gly Lys 
1               5                   10                  15      


<210>  14
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  isolated peptides analyzed by MS/MS


<220>
<221>  MOD_RES
<222>  (16)..(16)
<223>  may be ubiquitinated

<400>  14

Val Leu Gly Leu Ile Lys Pro Leu Glu Met Leu Gln Asp Gln Gly Lys 
1               5                   10                  15      


