                         SEQUENCE LISTING

<110>  Helmholtz Zentrum Mnchen Deutsches Forschungszentrum fr 
       Gesundheit
 
<120>  FNDC4 FUSION PROTEIN AND USES THEREOF

<130>  HEL17178PCT

<150>  LU 102453
<151>  2021-01-25

<160>  16   

<170>  PatentIn version 3.5

<210>  1
<211>  125
<212>  PRT
<213>  Homo Sapiens

<220>
<223>  Soluble Fibronectin type III domain-containing protein 4 

<400>  1

Asp Arg Pro Pro Ser Pro Val Asn Val Thr Val Thr His Leu Arg Ala 
1               5                   10                  15      


Asn Ser Ala Thr Val Ser Trp Asp Val Pro Glu Gly Asn Ile Val Ile 
            20                  25                  30          


Gly Tyr Ser Ile Ser Gln Gln Arg Gln Asn Gly Pro Gly Gln Arg Val 
        35                  40                  45              


Ile Arg Glu Val Asn Thr Thr Thr Arg Ala Cys Ala Leu Trp Gly Leu 
    50                  55                  60                  


Ala Glu Asp Ser Asp Tyr Thr Val Gln Val Arg Ser Ile Gly Leu Arg 
65                  70                  75                  80  


Gly Glu Ser Pro Pro Gly Pro Arg Val His Phe Arg Thr Leu Lys Gly 
                85                  90                  95      


Ser Asp Arg Leu Pro Ser Asn Ser Ser Ser Pro Gly Asp Ile Thr Val 
            100                 105                 110         


Glu Gly Leu Asp Gly Glu Arg Pro Leu Gln Thr Gly Glu 
        115                 120                 125 


<210>  2
<211>  125
<212>  PRT
<213>  Mus musculus

<220>
<223>  Soluble Fibronectin type III domain-containing protein 4 of 

<400>  2

Asp Arg Pro Pro Ser Pro Val Asn Val Thr Val Thr His Leu Arg Ala 
1               5                   10                  15      


Asn Ser Ala Thr Val Ser Trp Asp Val Pro Glu Gly Asn Ile Val Ile 
            20                  25                  30          


Gly Tyr Ser Ile Ser Gln Gln Arg Gln Asn Gly Pro Gly Gln Arg Val 
        35                  40                  45              


Ile Arg Glu Val Asn Thr Thr Thr Arg Ala Cys Ala Leu Trp Gly Leu 
    50                  55                  60                  


Ala Glu Asp Ser Asp Tyr Thr Val Gln Val Arg Ser Ile Gly Leu Arg 
65                  70                  75                  80  


Gly Glu Ser Pro Pro Gly Pro Arg Val His Phe Arg Thr Leu Lys Gly 
                85                  90                  95      


Ser Asp Arg Leu Pro Ser Asn Ser Ser Ser Pro Gly Asp Ile Thr Val 
            100                 105                 110         


Glu Gly Leu Asp Gly Glu Arg Pro Leu Gln Thr Gly Glu 
        115                 120                 125 


<210>  3
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Peptide Linker 

<220>
<221>  misc_feature
<222>  (8)..(9)
<223>  Xaa can be any naturally occurring amino acid

<400>  3

Glu Asn Leu Thr Phe Gln Gly Xaa Xaa 
1               5                   


<210>  4
<211>  381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  FcsFNDC4 comprising human soluble FNDC4

<400>  4

His His His His His His Ala Asp Leu Ile Glu Gly Arg Gly Asp Pro 
1               5                   10                  15      


Lys Ser Cys Asp Lys Pro His Thr Cys Pro Leu Cys Pro Ala Pro Glu 
            20                  25                  30          


Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 
        35                  40                  45              


Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 
    50                  55                  60                  


Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 
65                  70                  75                  80  


Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 
                85                  90                  95      


Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 
            100                 105                 110         


Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 
        115                 120                 125             


Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 
    130                 135                 140                 


Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 
145                 150                 155                 160 


Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 
                165                 170                 175     


Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Ala 
            180                 185                 190         


Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 
        195                 200                 205             


Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 
    210                 215                 220                 


Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 
225                 230                 235                 240 


Ser Leu Ser Pro Gly Lys Ala Ala Glu Asn Leu Thr Phe Gln Gly Ala 
                245                 250                 255     


Ala Arg Pro Pro Ser Pro Val Asn Val Thr Val Thr His Leu Arg Ala 
            260                 265                 270         


Asn Ser Ala Thr Val Ser Trp Asp Val Pro Glu Gly Asn Ile Val Ile 
        275                 280                 285             


Gly Tyr Ser Ile Ser Gln Gln Arg Gln Asn Gly Pro Gly Gln Arg Val 
    290                 295                 300                 


Ile Arg Glu Val Asn Thr Thr Thr Arg Ala Cys Ala Leu Trp Gly Leu 
305                 310                 315                 320 


Ala Glu Asp Ser Asp Tyr Thr Val Gln Val Arg Ser Ile Gly Leu Arg 
                325                 330                 335     


Gly Glu Ser Pro Pro Gly Pro Arg Val His Phe Arg Thr Leu Lys Gly 
            340                 345                 350         


Ser Asp Arg Leu Pro Ser Asn Ser Ser Ser Pro Gly Asp Ile Thr Val 
        355                 360                 365             


Glu Gly Leu Asp Gly Glu Arg Pro Leu Gln Thr Gly Glu 
    370                 375                 380     


<210>  5
<211>  454
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  FcsFNDC4 comprising mouse soluble FNDC4

<400>  5

His His His His His His Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 
1               5                   10                  15      


Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 
            20                  25                  30          


Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 
        35                  40                  45              


Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 
    50                  55                  60                  


Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Pro 
65                  70                  75                  80  


Arg Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 
                85                  90                  95      


Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 
            100                 105                 110         


Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 
        115                 120                 125             


Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 
    130                 135                 140                 


Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 
145                 150                 155                 160 


Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 
                165                 170                 175     


Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 
            180                 185                 190         


His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 
        195                 200                 205             


Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 
    210                 215                 220                 


Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 
225                 230                 235                 240 


Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 
                245                 250                 255     


Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 
            260                 265                 270         


Asn Tyr Lys Asn Thr Gln Pro Ile Met Asn Thr Asn Gly Ser Tyr Phe 
        275                 280                 285             


Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 
    290                 295                 300                 


Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 
305                 310                 315                 320 


Glu Lys Ser Leu Ser His Ser Pro Gly Lys Arg Pro Pro Ser Pro Val 
                325                 330                 335     


Asn Val Thr Val Thr His Leu Arg Ala Asn Ser Ala Thr Val Ser Trp 
            340                 345                 350         


Asp Val Pro Glu Gly Asn Ile Val Ile Gly Tyr Ser Ile Ser Gln Gln 
        355                 360                 365             


Arg Gln Asn Gly Pro Gly Gln Arg Val Ile Arg Glu Val Asn Thr Thr 
    370                 375                 380                 


Thr Arg Ala Cys Ala Leu Trp Gly Leu Ala Glu Asp Ser Asp Tyr Thr 
385                 390                 395                 400 


Val Gln Val Arg Ser Ile Gly Leu Arg Gly Glu Ser Pro Pro Gly Pro 
                405                 410                 415     


Arg Val His Phe Arg Thr Leu Lys Gly Ser Asp Arg Leu Pro Ser Asn 
            420                 425                 430         


Ser Ser Ser Pro Gly Asp Ile Thr Val Glu Gly Leu Asp Gly Glu Arg 
        435                 440                 445             


Pro Leu Gln Thr Gly Glu 
    450                 


<210>  6
<211>  242
<212>  PRT
<213>  Homo Sapiens

<400>  6

Ala Asp Leu Ile Glu Gly Arg Gly Asp Pro Lys Ser Cys Asp Lys Pro 
1               5                   10                  15      


His Thr Cys Pro Leu Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 
            20                  25                  30          


Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 
        35                  40                  45              


Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 
    50                  55                  60                  


Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 
65                  70                  75                  80  


Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 
                85                  90                  95      


Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 
            100                 105                 110         


Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 
        115                 120                 125             


Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 
    130                 135                 140                 


Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 
145                 150                 155                 160 


Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 
                165                 170                 175     


Asn Gly Gln Pro Glu Asn Asn Tyr Lys Ala Thr Pro Pro Val Leu Asp 
            180                 185                 190         


Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 
        195                 200                 205             


Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 
    210                 215                 220                 


Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
225                 230                 235                 240 


Ala Ala 
        


<210>  7
<211>  324
<212>  PRT
<213>  Mus Musculus

<400>  7

Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala 
1               5                   10                  15      


Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr 
            20                  25                  30          


Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser 
        35                  40                  45              


Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu 
    50                  55                  60                  


Ser Ser Ser Val Thr Val Pro Ser Ser Pro Arg Pro Ser Glu Thr Val 
65                  70                  75                  80  


Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys 
                85                  90                  95      


Ile Val Pro Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro 
            100                 105                 110         


Glu Val Ser Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu 
        115                 120                 125             


Thr Ile Thr Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ser 
    130                 135                 140                 


Lys Asp Asp Pro Glu Val Gln Phe Ser Trp Phe Val Asp Asp Val Glu 
145                 150                 155                 160 


Val His Thr Ala Gln Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr 
                165                 170                 175     


Phe Arg Ser Val Ser Glu Leu Pro Ile Met His Gln Asp Trp Leu Asn 
            180                 185                 190         


Gly Lys Glu Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro 
        195                 200                 205             


Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln 
    210                 215                 220                 


Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln Met Ala Lys Asp Lys Val 
225                 230                 235                 240 


Ser Leu Thr Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val 
                245                 250                 255     


Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln 
            260                 265                 270         


Pro Ile Met Asn Thr Asn Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn 
        275                 280                 285             


Val Gln Lys Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val 
    290                 295                 300                 


Leu His Glu Gly Leu His Asn His His Thr Glu Lys Ser Leu Ser His 
305                 310                 315                 320 


Ser Pro Gly Lys 
                


<210>  8
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TEV protease site

<220>
<221>  misc_feature
<222>  (4)..(4)
<223>  Xaa is tyrosine or threonine

<220>
<221>  misc_feature
<222>  (7)..(7)
<223>  Xaa is glycine or serine

<400>  8

Glu Asn Leu Xaa Phe Gln Xaa 
1               5                   


<210>  9
<211>  29
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Signal peptide

<400>  9

Met Pro Pro Gly Pro Cys Ala Trp Pro Pro Arg Ala Ala Leu Arg Leu 
1               5                   10                  15      


Trp Leu Gly Cys Val Cys Phe Ala Leu Val Gln Ala Asp 
            20                  25                  


<210>  10
<211>  31
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer to amplify extracellular part of FNDC4

<400>  10
gagagcggcc gctcgacctc cctctcctgt g                                      31


<210>  11
<211>  30
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer to amplify extracellular part of FNDC4

<400>  11
gagagaattc attcccctgt ctgcaatggc                                        30


<210>  12
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequences for silencing mouse Gpr116

<400>  12
cgtcatctta gacaagtctg ccttgaact                                         29


<210>  13
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequences for silencing mouse Gpr116

<400>  13
tactccattc acaccactgt catcaacaa                                         29


<210>  14
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequences for silencing mouse Gpr116

<400>  14
tcgcagtgtt ctgccacttc accaatgca                                        29


<210>  15
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequences for silencing mouse Gpr116

<400>  15
tgtggctggt gctatccacg acggtcgct                                        29


<210>  16
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequences for silencing mouse Gpr116

<400>  16
gcactaccag agctaactca gatagtact                                     	29