                         SEQUENCE LISTING

<110>  F. Hoffmann-La Roche AG
       Hoffmann-La Roche Inc.
 
<120>  Mammalian cell lines with SIRT-1 gene knockout

<130>  P35610-WO

<150>  EP19182558.7
<151>  2019-06-26

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  L3

<400>  1
ataacttcgt ataaagtctc ctatacgaag ttat                                   34


<210>  2
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  2L

<400>  2
ataacttcgt atagcataca ttatacgaag ttat                                   34


<210>  3
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  loxFas

<400>  3
acaacttcgt atataccttt ctatacgaag ttgt                                   34


<210>  4
<211>  608
<212>  DNA
<213>  Human cytomegalovirus

<400>  4
gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata       60

gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc      120

ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag      180

ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac      240

atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg      300

cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg      360

tattagtcat cgctattagc atggtgatgc ggttttggca gtacatcaat gggcgtggat      420

agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt      480

tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc      540

aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctccg tttagtgaac      600

gtcagatc                                                               608


<210>  5
<211>  696
<212>  DNA
<213>  Human cytomegalovirus

<400>  5
gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata       60

gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc      120

ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag      180

ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac      240

atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg      300

cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg      360

tattagtcat cgctattagc atggtgatgc ggttttggca gtacatcaat gggcgtggat      420

agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt      480

tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc      540

aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctccg tttagtgaac      600

gtcagatcta gctctgggag aggagcccag cactagaagt cggcggtgtt tccattcggt      660

gatcagcact gaacacagag gaagcttgcc gccacc                                696


<210>  6
<211>  2125
<212>  DNA
<213>  Human cytomegalovirus

<400>  6
ctgcagtgaa taataaaatg tgtgtttgtc cgaaatacgc gttttgagat ttctgtcgcc       60

gactaaattc atgtcgcgcg atagtggtgt ttatcgccga tagagatggc gatattggaa      120

aaatcgatat ttgaaaatat ggcatattga aaatgtcgcc gatgtgagtt tctgtgtaac      180

tgatatcgcc atttttccaa aagtgatttt tgggcatacg cgatatctgg cgatagcgct      240

tatatcgttt acgggggatg gcgatagacg actttggtga cttgggcgat tctgtgtgtc      300

gcaaatatcg cagtttcgat ataggtgaca gacgatatga ggctatatcg ccgatagagg      360

cgacatcaag ctggcacatg gccaatgcat atcgatctat acattgaatc aatattggcc      420

attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca      480

tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc      540

atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca      600

tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc      660

gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat      720

agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt      780

acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc      840

cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta      900

cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg      960

atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt     1020

gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac     1080

gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa     1140

ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga     1200

ccgatccagc ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag     1260

tgacgtaagt accgcctata gagtctatag gcccaccccc ttggcttctt atgcatgcta     1320

tactgttttt ggcttggggt ctatacaccc ccgcttcctc atgttatagg tgatggtata     1380

gcttagccta taggtgtggg ttattgacca ttattgacca ctcccctatt ggtgacgata     1440

ctttccatta ctaatccata acatggctct ttgccacaac tctctttatt ggctatatgc     1500

caatacactg tccttcagag actgacacgg actctgtatt tttacaggat ggggtctcat     1560

ttattattta caaattcaca tatacaacac caccgtcccc agtgcccgca gtttttatta     1620

aacataacgt gggatctcca cgcgaatctc gggtacgtgt tccggacatg ggctcttctc     1680

cggtagcggc ggagcttcta catccgagcc ctgctcccat gcctccagcg actcatggtc     1740

gctcggcagc tccttgctcc taacagtgga ggccagactt aggcacagca cgatgcccac     1800

caccaccagt gtgccgcaca aggccgtggc ggtagggtat gtgtctgaaa atgagctcgg     1860

ggagcgggct tgcaccgctg acgcatttgg aagacttaag gcagcggcag aagaagatgc     1920

aggcagctga gttgttgtgt tctgataaga gtcagaggta actcccgttg cggtgctgtt     1980

aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg ccaccagaca     2040

taatagctga cagactaaca gactgttcct ttccatgggt cttttctgca gtcaccgtcc     2100

ttgacacggt ttaaacgccg ccacc                                           2125


<210>  7
<211>  129
<212>  DNA
<213>  Simian virus 40

<400>  7
aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca       60

aataaagcat ttttttcacc attctagttg tggtttgtcc aaactcatca atgtatctta      120

tcatgtctg                                                              129


<210>  8
<211>  225
<212>  DNA
<213>  Bos taurus

<400>  8
ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc       60

tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc      120

tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt      180

gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgg                      225


<210>  9
<211>  73
<212>  DNA
<213>  Homo sapiens

<400>  9
caggataata tatggtaggg ttcatagcca gagtaacctt tttttttaat ttttatttta       60

ttttattttt gag                                                          73


<210>  10
<211>  288
<212>  DNA
<213>  Simian virus 40

<400>  10
agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca       60

tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa      120

ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag      180

aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag      240

gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcg                   288


<210>  11
<211>  798
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  green fluorescent protein encoding nucleic acid

<400>  11
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac       60

ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac      120

ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc      180

ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag      240

cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc      300

ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg      360

gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac      420

aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac      480

ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc      540

gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac      600

tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc      660

ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc      720

ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga      780

tccaccggat ctagatga                                                    798


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gRNA_SIRT1_1

<400>  12
tatcatccaa ctcaggtgga                                                   20


<210>  13
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gRNA_SIRT1_2

<400>  13
gcagcatctc atgattggca                                                   20


<210>  14
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gRNA_SIRT1_3

<400>  14
gcattcttga agtaacttca                                                   20


<210>  15
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  oSA060_SIRT1_for

<400>  15
gctgcccttc aagttatggc                                                   20


<210>  16
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  oSA061_SIRT1_rev

<400>  16
gctggccttt tgactcacag                                                   20


<210>  17
<211>  747
<212>  PRT
<213>  Homo sapiens

<400>  17

Met Ala Asp Glu Ala Ala Leu Ala Leu Gln Pro Gly Gly Ser Pro Ser 
1               5                   10                  15      


Ala Ala Gly Ala Asp Arg Glu Ala Ala Ser Ser Pro Ala Gly Glu Pro 
            20                  25                  30          


Leu Arg Lys Arg Pro Arg Arg Asp Gly Pro Gly Leu Glu Arg Ser Pro 
        35                  40                  45              


Gly Glu Pro Gly Gly Ala Ala Pro Glu Arg Glu Val Pro Ala Ala Ala 
    50                  55                  60                  


Arg Gly Cys Pro Gly Ala Ala Ala Ala Ala Leu Trp Arg Glu Ala Glu 
65                  70                  75                  80  


Ala Glu Ala Ala Ala Ala Gly Gly Glu Gln Glu Ala Gln Ala Thr Ala 
                85                  90                  95      


Ala Ala Gly Glu Gly Asp Asn Gly Pro Gly Leu Gln Gly Pro Ser Arg 
            100                 105                 110         


Glu Pro Pro Leu Ala Asp Asn Leu Tyr Asp Glu Asp Asp Asp Asp Glu 
        115                 120                 125             


Gly Glu Glu Glu Glu Glu Ala Ala Ala Ala Ala Ile Gly Tyr Arg Asp 
    130                 135                 140                 


Asn Leu Leu Phe Gly Asp Glu Ile Ile Thr Asn Gly Phe His Ser Cys 
145                 150                 155                 160 


Glu Ser Asp Glu Glu Asp Arg Ala Ser His Ala Ser Ser Ser Asp Trp 
                165                 170                 175     


Thr Pro Arg Pro Arg Ile Gly Pro Tyr Thr Phe Val Gln Gln His Leu 
            180                 185                 190         


Met Ile Gly Thr Asp Pro Arg Thr Ile Leu Lys Asp Leu Leu Pro Glu 
        195                 200                 205             


Thr Ile Pro Pro Pro Glu Leu Asp Asp Met Thr Leu Trp Gln Ile Val 
    210                 215                 220                 


Ile Asn Ile Leu Ser Glu Pro Pro Lys Arg Lys Lys Arg Lys Asp Ile 
225                 230                 235                 240 


Asn Thr Ile Glu Asp Ala Val Lys Leu Leu Gln Glu Cys Lys Lys Ile 
                245                 250                 255     


Ile Val Leu Thr Gly Ala Gly Val Ser Val Ser Cys Gly Ile Pro Asp 
            260                 265                 270         


Phe Arg Ser Arg Asp Gly Ile Tyr Ala Arg Leu Ala Val Asp Phe Pro 
        275                 280                 285             


Asp Leu Pro Asp Pro Gln Ala Met Phe Asp Ile Glu Tyr Phe Arg Lys 
    290                 295                 300                 


Asp Pro Arg Pro Phe Phe Lys Phe Ala Lys Glu Ile Tyr Pro Gly Gln 
305                 310                 315                 320 


Phe Gln Pro Ser Leu Cys His Lys Phe Ile Ala Leu Ser Asp Lys Glu 
                325                 330                 335     


Gly Lys Leu Leu Arg Asn Tyr Thr Gln Asn Ile Asp Thr Leu Glu Gln 
            340                 345                 350         


Val Ala Gly Ile Gln Arg Ile Ile Gln Cys His Gly Ser Phe Ala Thr 
        355                 360                 365             


Ala Ser Cys Leu Ile Cys Lys Tyr Lys Val Asp Cys Glu Ala Val Arg 
    370                 375                 380                 


Gly Asp Ile Phe Asn Gln Val Val Pro Arg Cys Pro Arg Cys Pro Ala 
385                 390                 395                 400 


Asp Glu Pro Leu Ala Ile Met Lys Pro Glu Ile Val Phe Phe Gly Glu 
                405                 410                 415     


Asn Leu Pro Glu Gln Phe His Arg Ala Met Lys Tyr Asp Lys Asp Glu 
            420                 425                 430         


Val Asp Leu Leu Ile Val Ile Gly Ser Ser Leu Lys Val Arg Pro Val 
        435                 440                 445             


Ala Leu Ile Pro Ser Ser Ile Pro His Glu Val Pro Gln Ile Leu Ile 
    450                 455                 460                 


Asn Arg Glu Pro Leu Pro His Leu His Phe Asp Val Glu Leu Leu Gly 
465                 470                 475                 480 


Asp Cys Asp Val Ile Ile Asn Glu Leu Cys His Arg Leu Gly Gly Glu 
                485                 490                 495     


Tyr Ala Lys Leu Cys Cys Asn Pro Val Lys Leu Ser Glu Ile Thr Glu 
            500                 505                 510         


Lys Pro Pro Arg Thr Gln Lys Glu Leu Ala Tyr Leu Ser Glu Leu Pro 
        515                 520                 525             


Pro Thr Pro Leu His Val Ser Glu Asp Ser Ser Ser Pro Glu Arg Thr 
    530                 535                 540                 


Ser Pro Pro Asp Ser Ser Val Ile Val Thr Leu Leu Asp Gln Ala Ala 
545                 550                 555                 560 


Lys Ser Asn Asp Asp Leu Asp Val Ser Glu Ser Lys Gly Cys Met Glu 
                565                 570                 575     


Glu Lys Pro Gln Glu Val Gln Thr Ser Arg Asn Val Glu Ser Ile Ala 
            580                 585                 590         


Glu Gln Met Glu Asn Pro Asp Leu Lys Asn Val Gly Ser Ser Thr Gly 
        595                 600                 605             


Glu Lys Asn Glu Arg Thr Ser Val Ala Gly Thr Val Arg Lys Cys Trp 
    610                 615                 620                 


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


Gln Tyr Leu Phe Leu Pro Pro Asn Arg Tyr Ile Phe His Gly Ala Glu 
                645                 650                 655     


Val Tyr Ser Asp Ser Glu Asp Asp Val Leu Ser Ser Ser Ser Cys Gly 
            660                 665                 670         


Ser Asn Ser Asp Ser Gly Thr Cys Gln Ser Pro Ser Leu Glu Glu Pro 
        675                 680                 685             


Met Glu Asp Glu Ser Glu Ile Glu Glu Phe Tyr Asn Gly Leu Glu Asp 
    690                 695                 700                 


Glu Pro Asp Val Pro Glu Arg Ala Gly Gly Ala Gly Phe Gly Thr Asp 
705                 710                 715                 720 


Gly Asp Asp Gln Glu Ala Ile Asn Glu Ala Ile Ser Val Lys Gln Glu 
                725                 730                 735     


Val Thr Asp Met Asn Tyr Pro Ser Asn Lys Ser 
            740                 745         


<210>  18
<211>  693
<212>  PRT
<213>  Cricetulus griseus

<400>  18

Met Glu Ala Ala Ser Gln Pro Ala Asp Glu Pro Leu Arg Lys Ser Ala 
1               5                   10                  15      


Gln Arg Arg Ala Trp Pro Arg Ala Gln Pro Gly Arg Val Glu Pro Gly 
            20                  25                  30          


Gly Arg Gly Gly Gly Gly Gly Ala Ala Ala Ala Ala Ala Gly Cys Glu 
        35                  40                  45              


Ala Ala Ser Ala Ala Ala Pro Ala Ala Ala Leu Trp Arg Glu Ala Val 
    50                  55                  60                  


Gly Ala Ala Ala Ala Gly Ala Glu Gln Glu Ala Arg Arg Pro Pro Arg 
65                  70                  75                  80  


Pro Gly Arg Lys Gln Trp Val Gly Pro Ala Ala Gly Ala Glu Gly Cys 
                85                  90                  95      


Arg Arg Leu Arg Arg Arg Arg Gly Arg Gly Gly Arg Arg Gly Gly Gly 
            100                 105                 110         


Gly Gly Gly Gly Gly Ser Asp Arg Leu Pro Arg Phe Ser Phe Ser Gly 
        115                 120                 125             


Ile Gly Pro Tyr Thr Phe Val Gln Gln His Leu Met Ile Gly Thr Asp 
    130                 135                 140                 


Pro Arg Thr Ile Leu Lys Asp Leu Leu Pro Glu Thr Ile Pro Pro Pro 
145                 150                 155                 160 


Glu Leu Asp Asp Met Thr Leu Trp Gln Ile Val Ile Asn Ile Leu Ser 
                165                 170                 175     


Glu Pro Pro Lys Arg Lys Lys Arg Lys Asp Ile Asn Thr Ile Glu Asp 
            180                 185                 190         


Ala Val Lys Leu Leu Gln Glu Cys Lys Lys Ile Ile Val Leu Thr Gly 
        195                 200                 205             


Ala Gly Val Ser Val Ser Cys Gly Ile Pro Asp Phe Arg Ser Arg Asp 
    210                 215                 220                 


Gly Ile Tyr Ala Arg Leu Ala Val Asp Phe Pro Asp Leu Pro Asp Pro 
225                 230                 235                 240 


Gln Ala Met Phe Asp Ile Glu Tyr Phe Arg Lys Asp Pro Arg Pro Phe 
                245                 250                 255     


Phe Lys Phe Ala Lys Glu Ile Tyr Pro Gly Gln Phe Gln Pro Ser Leu 
            260                 265                 270         


Cys His Lys Phe Ile Ala Leu Ser Asp Lys Glu Gly Lys Leu Leu Arg 
        275                 280                 285             


Asn Tyr Thr Gln Asn Ile Asp Thr Leu Glu Gln Val Ala Gly Ile Gln 
    290                 295                 300                 


Arg Ile Ile Gln Cys His Gly Ser Phe Ala Thr Ala Ser Cys Leu Ile 
305                 310                 315                 320 


Cys Lys Tyr Lys Val Asp Cys Glu Thr Val Arg Gly Asp Ile Phe Asn 
                325                 330                 335     


Gln Val Val Pro Arg Cys Pro Arg Cys Pro Ala Asp Glu Pro Leu Ala 
            340                 345                 350         


Ile Met Lys Pro Glu Ile Val Phe Phe Gly Glu Asn Leu Pro Glu Gln 
        355                 360                 365             


Phe His Arg Ala Met Lys Tyr Asp Lys Asp Glu Val Asp Leu Leu Ile 
    370                 375                 380                 


Val Ile Gly Ser Ser Leu Lys Val Arg Pro Val Ala Leu Ile Pro Ser 
385                 390                 395                 400 


Ser Ile Pro His Glu Val Pro Gln Ile Leu Ile Asn Arg Glu Pro Leu 
                405                 410                 415     


Pro His Leu His Phe Asp Val Glu Leu Leu Gly Asp Cys Asp Val Ile 
            420                 425                 430         


Ile Asn Glu Leu Cys His Arg Leu Gly Gly Glu Tyr Ala Lys Leu Cys 
        435                 440                 445             


Cys Asn Pro Val Lys Leu Ser Glu Ile Thr Glu Lys Pro Pro Arg Thr 
    450                 455                 460                 


Gln Lys Glu Leu Val His Leu Ser Glu Leu Pro Pro Thr Pro Leu His 
465                 470                 475                 480 


Ile Ser Glu Asp Ser Ser Ser Pro Glu Arg Thr Val Pro Gln Asp Ser 
                485                 490                 495     


Ala Ser Met Ile Ala Thr Cys Val Asp Gln Ala Thr Glu Asn Arg Val 
            500                 505                 510         


Asp Leu Glu Val Ser Glu Ser Glu Gly Cys Val Glu Glu Lys Ser Gln 
        515                 520                 525             


Glu Ala Gln Thr Cys Arg Ser Val Glu Ser Ile Asn Val Glu Asn Pro 
    530                 535                 540                 


Asp Leu Lys Ala Val Gly Ser Ser Thr Gly Glu Lys Asn Glu Arg Thr 
545                 550                 555                 560 


Ser Val Ala Glu Ala Val Arg Lys Cys Trp Pro Asn Arg Leu Ala Lys 
                565                 570                 575     


Glu Gln Ile Ser Lys Arg Leu Asp Gly Asn Gln Tyr Leu Phe Val Pro 
            580                 585                 590         


Pro Asn Arg Tyr Ile Phe His Gly Ala Glu Val Tyr Ser Asp Ser Glu 
        595                 600                 605             


Asp Asp Ile Leu Ser Ser Ser Ser Cys Gly Ser Asn Ser Asp Ser Gly 
    610                 615                 620                 


Thr Cys Gln Ser Pro Ser Leu Glu Glu Pro Met Glu Asp Glu Ser Glu 
625                 630                 635                 640 


Ile Glu Glu Phe Tyr Asn Gly Leu Glu Asp Asp Thr Asp Arg Pro Glu 
                645                 650                 655     


Gly Thr Gly Gly Ser Gly Leu Gly Val Asp Gly Gly Glu Gln Glu Leu 
            660                 665                 670         


Val His Glu Ala Thr Ala Met Arg Gln Glu Ala Thr Asp Val Asn Tyr 
        675                 680                 685             


Pro Ser Asp Lys Ser 
    690             


