                         SEQUENCE LISTING

<110>  Inserm
 
<120>  BIFUNCTIONAL VECTORS ALLOWING BCL11A SILENCING AND EXPRESSION OF 
       AN ANTI-SICKLING HBB AND USES THEREOF FOR GENE THERAPY OF 
       B-HEMOGLOBINOPATHIES

<130>  BIO19013 MICCIO / MC

<160>  28    

<170>  PatentIn version 3.3

<210>  1
<211>  257
<212>  DNA
<213>  Artificial

<220>
<223>  bAS3 intron 2 sequence (5'-3')

<400>  1
gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt     60

cataggaagg ggagaagtaa cagggtattt ctgcatataa attgtaactg atgtaagagg    120

tttcatattg ctaatagcag ctacaatcca gctaccattc tgcttttatt ttatggttgg    180

gataaggctg gattattctg agtccaagct aggccctttt gctaatcatg ttcatacctc    240

ttatcttcct cccacag                                                   257


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

<220>
<223>  guide strand-shRNA BCL11A-XL

<400>  2
gcgcgatcga gtgttgaata a                                               21


<210>  3
<211>  15
<212>  DNA
<213>  Artificial

<220>
<223>  loop segment

<400>  3
ctccatgtgg tagag                                                      15


<210>  4
<211>  57
<212>  DNA
<213>  Artificial

<220>
<223>  shRNA BCL11A-XL

<400>  4
gcgcgatcga gtgttgaata actccatgtg gtagagttat tcaacactcg atcgcgc        57


<210>  5
<211>  104
<212>  DNA
<213>  Artificial

<220>
<223>  amiR-shRNA BCL11A-XL

<400>  5
cctggcctcc tgcagtgcca cgctgcgcga tcgagtgttg aataactcca tgtggtagag     60

ttattcaaca ctcgatcgcg cagtgcggca catgcttacc agct                     104


<210>  6
<211>  361
<212>  DNA
<213>  Artificial

<220>
<223>  (bAS3-miR/int2_del/amiR-shRNA BCL11A-XL

<400>  6
gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt     60

cataggaagg ggagaagtaa cagggcctgg cctcctgcag tgccacgctg cgcgatcgag    120

tgttgaataa ctccatgtgg tagagttatt caacactcga tcgcgcagtg cggcacatgc    180

ttaccagctt atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata    240

gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag gctggattat    300

tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca    360

g                                                                    361


<210>  7
<211>  361
<212>  DNA
<213>  Artificial

<220>
<223>  bAS3-miR/int2/amiR-shRNA BCL11A-XL

<400>  7
gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt     60

cataggaagg ggagaagtaa cagggtattt ctgcatataa attgtaactg atgtaagagg    120

tttcatattg ctaatagcag ctacaacctg gcctcctgca gtgccacgct gcgcgatcga    180

gtgttgaata actccatgtg gtagagttat tcaacactcg atcgcgcagt gcggcacatg    240

cttaccagct tccagctacc attctgcttt tattttatgg ttgggataag gctggattat    300

tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca    360

g                                                                    361


<210>  8
<211>  147
<212>  PRT
<213>  Homo sapiens

<400>  8

Met Val His Leu Thr Pro Glu Glu Lys Ser Ala Val Thr Ala Leu Trp 
1               5                   10                  15      


Gly Lys Val Asn Val Asp Glu Val Gly Gly Glu Ala Leu Gly Arg Leu 
            20                  25                  30          


Leu Val Val Tyr Pro Trp Thr Gln Arg Phe Phe Glu Ser Phe Gly Asp 
        35                  40                  45              


Leu Ser Thr Pro Asp Ala Val Met Gly Asn Pro Lys Val Lys Ala His 
    50                  55                  60                  


Gly Lys Lys Val Leu Gly Ala Phe Ser Asp Gly Leu Ala His Leu Asp 
65                  70                  75                  80  


Asn Leu Lys Gly Thr Phe Ala Thr Leu Ser Glu Leu His Cys Asp Lys 
                85                  90                  95      


Leu His Val Asp Pro Glu Asn Phe Arg Leu Leu Gly Asn Val Leu Val 
            100                 105                 110         


Cys Val Leu Ala His His Phe Gly Lys Glu Phe Thr Pro Pro Val Gln 
        115                 120                 125             


Ala Ala Tyr Gln Lys Val Val Ala Gly Val Ala Asn Ala Leu Ala His 
    130                 135                 140                 


Lys Tyr His 
145         


<210>  9
<211>  2051
<212>  DNA
<213>  Artificial

<220>
<223>  > bAS3 sequence (5'-3') + ( bAS3-miR/int2_del/amiR-shRNA 
       BCL11A-XL)

<400>  9
acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc atggtgcacc     60

tgactcctga ggagaagtct gccgttactg ccctgtggga caaggtgaac gtggatgccg    120

ttggtggtga ggccctgggc aggttggtat caaggttaca agacaggttt aaggagacca    180

atagaaactg ggcatgtgga gacagagaag actcttgggt ttctgatagg cactgactct    240

ctctgcctat tggtctattt tcccaccctt aggctgctgg tggtctaccc ttggacccag    300

aggttctttg agtcctttgg ggatctgtcc actcctgatg ctgttatggg caaccctaag    360

gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc tcacctggac    420

aacctcaagg gcacctttgc ccagctgagt gagctgcact gtgacaagct gcacgtggat    480

cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta    540

tggttaagtt catgtcatag gaaggggaga agtaacaggg cctggcctcc tgcagtgcca    600

cgctgcgcga tcgagtgttg aataactcca tgtggtagag ttattcaaca ctcgatcgcg    660

cagtgcggca catgcttacc agcttatttc tgcatataaa ttgtaactga tgtaagaggt    720

ttcatattgc taatagcagc tacaatccag ctaccattct gcttttattt tatggttggg    780

ataaggctgg attattctga gtccaagcta ggcccttttg ctaatcatgt tcatacctct    840

tatcttcctc ccacagctcc tgggcaacgt gctggtctgt gtgctggccc atcactttgg    900

caaagaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg gtgtggctaa    960

tgccctggcc cacaagtatc actaagctcg ctttcttgct gtccaatttc tattaaaggt   1020

tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc cttgagcatc   1080

tggattctgc ctaataaaaa acatttattt tcattgcaat gatgtattta aattatttct   1140

gaatatttta ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa   1200

gagctagttc aaaccttggg aaaatacact atatcttaaa ctccatgaaa gaaggtgagg   1260

ctgcaaacag ctaatgcaca ttggcaacag cccctgatgc ctatgcctta ttcatccctc   1320

agaaaaggat tcaagtagag gcttgatttg gaggttaaag ttttgctatg ctgtatttta   1380

cattacttat tgttttagct gtcctcatgg tacgtaccga taaaattttg aattttgtaa   1440

tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact attctttccc   1500

ctgcactgta ccccccaatc cccccttttc ttttaaaagt taaccgatac cgtcgagatc   1560

cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct tctattcctt   1620

cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc   1680

cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc   1740

ctcctactat cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat   1800

tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt   1860

tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc   1920

actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat   1980

tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct   2040

gttgatcctt t                                                        2051


<210>  10
<211>  2051
<212>  DNA
<213>  Artificial

<220>
<223>  bAS3 sequence (5'-3') + (bAS3-miR/int2/amiR-shRNA BCL11A-XL)

<400>  10
acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc atggtgcacc     60

tgactcctga ggagaagtct gccgttactg ccctgtggga caaggtgaac gtggatgccg    120

ttggtggtga ggccctgggc aggttggtat caaggttaca agacaggttt aaggagacca    180

atagaaactg ggcatgtgga gacagagaag actcttgggt ttctgatagg cactgactct    240

ctctgcctat tggtctattt tcccaccctt aggctgctgg tggtctaccc ttggacccag    300

aggttctttg agtcctttgg ggatctgtcc actcctgatg ctgttatggg caaccctaag    360

gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc tcacctggac    420

aacctcaagg gcacctttgc ccagctgagt gagctgcact gtgacaagct gcacgtggat    480

cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta    540

tggttaagtt catgtcatag gaaggggaga agtaacaggg tatttctgca tataaattgt    600

aactgatgta agaggtttca tattgctaat agcagctaca acctggcctc ctgcagtgcc    660

acgctgcgcg atcgagtgtt gaataactcc atgtggtaga gttattcaac actcgatcgc    720

gcagtgcggc acatgcttac cagcttccag ctaccattct gcttttattt tatggttggg    780

ataaggctgg attattctga gtccaagcta ggcccttttg ctaatcatgt tcatacctct    840

tatcttcctc ccacagctcc tgggcaacgt gctggtctgt gtgctggccc atcactttgg    900

caaagaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg gtgtggctaa    960

tgccctggcc cacaagtatc actaagctcg ctttcttgct gtccaatttc tattaaaggt   1020

tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc cttgagcatc   1080

tggattctgc ctaataaaaa acatttattt tcattgcaat gatgtattta aattatttct   1140

gaatatttta ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa   1200

gagctagttc aaaccttggg aaaatacact atatcttaaa ctccatgaaa gaaggtgagg   1260

ctgcaaacag ctaatgcaca ttggcaacag cccctgatgc ctatgcctta ttcatccctc   1320

agaaaaggat tcaagtagag gcttgatttg gaggttaaag ttttgctatg ctgtatttta   1380

cattacttat tgttttagct gtcctcatgg tacgtaccga taaaattttg aattttgtaa   1440

tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact attctttccc   1500

ctgcactgta ccccccaatc cccccttttc ttttaaaagt taaccgatac cgtcgagatc   1560

cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct tctattcctt   1620

cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc   1680

cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc   1740

ctcctactat cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat   1800

tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt   1860

tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc   1920

actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat   1980

tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct   2040

gttgatcctt t                                                        2051


<210>  11
<211>  26
<212>  DNA
<213>  Artificial

<220>
<223>  probe

<400>  11
cctgtcatgc ccacacaaat ctctcc                                          26


<210>  12
<211>  22
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  12
gctgtcatct cttgtgggct gt                                              22


<210>  13
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  13
actcatggga gctgctggtt c                                               21


<210>  14
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  probe

<400>  14
cgcacggcaa gaggcgagg                                                  19


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

<220>
<223>  primer

<400>  15
tcccccgctt aatactgacg                                                 20


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

<220>
<223>  primer

<400>  16
caggactcgg cttgctgaag                                                 20


<210>  17
<211>  22
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  17
gccaccactt tctgataggc ag                                              22


<210>  18
<211>  18
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  18
aagggcacct ttgcccag                                                   18


<210>  19
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  19
atgcgagctg tgcaactatg                                                 20


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

<220>
<223>  primer

<400>  20
gtaaacgtcc ttccccacct                                                 20


<210>  21
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  21
cctgtcctct gcctctgcc                                                  19


<210>  22
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  22
ggattgccaa aacggtcac                                                  19


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

<220>
<223>  primer

<400>  23
agttcacgcc caagtacatc                                                 20


<210>  24
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  24
cttcacagtc ctcatggcc                                                  19


<210>  25
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  primer

<400>  25
cggtcaactt caagctccta a                                               21


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

<220>
<223>  primer

<400>  26
acagaagcca ggaacttgtc                                                 20


<210>  27
<211>  104
<212>  RNA
<213>  Artificial

<220>
<223>  amiR sequence of Figure 1A


<220>
<221>  misc_feature
<222>  (25)..(45)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (61)..(81)
<223>  n is a, c, g, or u

<400>  27
ccuggccucc ugcagugcca cgcunnnnnn nnnnnnnnnn nnnnncucca ugugguagag     60

nnnnnnnnnn nnnnnnnnnn nagugcggca caugcuuacc agcu                     104


<210>  28
<211>  104
<212>  RNA
<213>  Artificial

<220>
<223>  amiR sequence of Figure 1B

<400>  28
ccuggccucc ugcagugcca cgcugcgcga ucgaguguug aauaacucca ugugguagag     60

uuauucaaca cucgaucgcg cagugcggca caugcuuacc agcu                     104


