                         SEQUENCE LISTING

<110>  President and Fellows of Harvard College
 
<120>  RNA-Guided Human Genome Engineering

<130>  010498.00464

<140>  PCT/US2013/075326
<141>  2013-12-16

<150>  US 61/779,169
<151>  2013-03-16

<150>  US 61/738,355
<151>  2012-12-17

<160>  38    

<170>  PatentIn version 3.5

<210>  1
<211>  268
<212>  DNA
<213>  Artificial

<220>
<223>  reference sequence

<400>  1
cacttcagga cagcatgttt gctgcctcca gggatcctgt gtccccgagc tgggaccacc       60

ttatattccc agggccggtt aatgtggctc tggttctggg tacttttatc tgtcccctcc      120

accccacagt ggggccacta gggacaggat tggtgacaga aaagccccat ccttaggcct      180

cctccttcct agtctcctga tattgggtct aacccccacc tcctgttagg cagattcctt      240

atctggtgac acacccccat ttcctgga                                         268


<210>  2
<211>  53
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  2
ctcggcattc ctgctgaacc gctcttccga tctacaggag gtgggggtta gac              53


<210>  3
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  3
acactctttc cctacacgac gctcttccga tctcgtgatt atattcccag ggccggtta        59


<210>  4
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  4
acactctttc cctacacgac gctcttccga tctacatcgt atattcccag ggccggtta        59


<210>  5
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  5
acactctttc cctacacgac gctcttccga tctgcctaat atattcccag ggccggtta        59


<210>  6
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  6
acactctttc cctacacgac gctcttccga tcttggtcat atattcccag ggccggtta        59


<210>  7
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  7
acactctttc cctacacgac gctcttccga tctcactgtt atattcccag ggccggtta        59


<210>  8
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  8
acactctttc cctacacgac gctcttccga tctattggct atattcccag ggccggtta        59


<210>  9
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  9
acactctttc cctacacgac gctcttccga tctgatctgt atattcccag ggccggtta        59


<210>  10
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  10
acactctttc cctacacgac gctcttccga tcttcaagtt atattcccag ggccggtta        59


<210>  11
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  11
acactctttc cctacacgac gctcttccga tctctgatct atattcccag ggccggtta        59


<210>  12
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  12
acactctttc cctacacgac gctcttccga tctaagctat atattcccag ggccggtta        59


<210>  13
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  13
acactctttc cctacacgac gctcttccga tctgtagcct atattcccag ggccggtta        59


<210>  14
<211>  59
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  14
acactctttc cctacacgac gctcttccga tcttacaagt atattcccag ggccggtta        59


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

<220>
<223>  PCR primer

<400>  15
ctgccgtctc tctcctgagt                                                   20


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

<220>
<223>  PCR primer

<400>  16
gtgggcttgt actcggtcat                                                   20


<210>  17
<211>  81
<212>  RNA
<213>  Artificial

<220>
<223>  guide RNA

<400>  17
guuuuagagc uagaaaauag caaguuaaaa uaaggcuagu ccguuaucaa cuugaaaaag       60

uggcaccgag ucggugcuuu u                                                 81


<210>  18
<211>  51
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  18
ttatctgtcc cctccacccc acagtggggc cactagggac aggattggtg a                51


<210>  19
<211>  4146
<212>  DNA
<213>  Artificial

<220>
<223>  Human codon optimized Cas9

<400>  19
gccaccatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggctgg       60

gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc      120

gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg      180

gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg      240

atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc      300

cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc      360

tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg      420

aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg      480

gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac      540

agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag      600

aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa      660

tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt      720

ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg      780

gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg      840

ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac      900

gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc      960

gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt     1020

gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc     1080

tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc     1140

atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg     1200

ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa     1260

ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa     1320

aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga     1380

aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc     1440

gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt     1500

gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc     1560

acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca     1620

ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa     1680

gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt     1740

gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg     1800

aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac     1860

attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact     1920

tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga     1980

tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca     2040

atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat     2100

gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac     2160

agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg     2220

cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat     2280

atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg     2340

gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa     2400

cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac     2460

ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg     2520

gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca     2580

agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa     2640

atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat     2700

aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa     2760

aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc     2820

atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg     2880

aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc     2940

aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc     3000

aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt     3060

aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt     3120

tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg     3180

aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg     3240

gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc     3300

gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag     3360

ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca     3420

gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa     3480

agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc     3540

atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt     3600

cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc     3660

gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg     3720

gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc     3780

gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa     3840

agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg     3900

gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg     3960

ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct     4020

acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca     4080

agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag     4140

gtgtga                                                                4146


<210>  20
<211>  455
<212>  DNA
<213>  Artificial

<220>
<223>  Guide oligonucleotide


<220>
<221>  misc_feature
<222>  (320)..(338)
<223>  wherein n is g, a, t or c

<400>  20
tgtacaaaaa agcaggcttt aaaggaacca attcagtcga ctggatccgg taccaaggtc       60

gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct      120

gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg      180

tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg      240

gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg      300

tggaaaggac gaaacaccgn nnnnnnnnnn nnnnnnnngt tttagagcta gaaatagcaa      360

gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt      420

tctagaccca gctttcttgt acaaagttgg catta                                 455


<210>  21
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  21
gtgaaccgca tcgagctgaa ggg                                               23


<210>  22
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  22
ggagcgcacc atcttcttca agg                                               23


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

<220>
<223>  target oligonucleotide

<400>  23
gtcccctcca ccccacagtg ggg                                               23


<210>  24
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  24
ggggccacta gggacaggat tgg                                               23


<210>  25
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  25
gcatgatgcg cggcccaagg agg                                               23


<210>  26
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  26
gagatgatcg ccccttcttc tgg                                               23


<210>  27
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  27
gaattactca cgccccaagg agg                                               23


<210>  28
<211>  51
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  28
ttatctgtcc cctccacccc acagtggggc cactagggac aggattggtg a                51


<210>  29
<211>  72
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  29
taatactttt atctgtcccc ctccacccca cagtggggcc actagggaca ggattggtga       60

cagaaaagcc cc                                                           72


<210>  30
<211>  70
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  30
taatacttta tcthtcaaaa aaaccccaca gtggggccac tagggacagg attggtgaca       60

gaaaagcccc                                                              70


<210>  31
<211>  71
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  31
taatactttt atctgtcggg gggaccccac atgttttcca ctagggacag gattggtgac       60

agaaaagccc c                                                            71


<210>  32
<211>  62
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  32
taaatgcatc atgcgcggcc caaggaggga gatgatcgcc ccttcttctg gctctttgag       60

aa                                                                      62


<210>  33
<211>  61
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  33
taatgaatta ctcacgcccc aaggagggtg atgaccggcc gttcttctgg atgtttgaga       60

a                                                                       61


<210>  34
<211>  109
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  34
ggtactttta tctgtcccct ccaccccaca gtggggccac tagggacagg attggtgaca       60

gaaaagcccc atccttaggc ctcctccttc ctagtctcct atattggtc                  109


<210>  35
<211>  90
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  35
ttatctgtcc cctccacccc acagtggggc cactagggac aggaaaggtg acagaaaagc       60

cccatcctta ggcctcctcc ttcctagtct                                        90


<210>  36
<211>  199
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide


<220>
<221>  misc_feature
<222>  (26)..(50)
<223>  wherein n is g, a, t or c

<220>
<221>  misc_feature
<222>  (91)..(109)
<223>  wherein n is g, a, t or c

<220>
<221>  misc_feature
<222>  (150)..(174)
<223>  wherein n is g, a, t or c

<400>  36
tatgaggacg aatctcccgc ttatannnnn nnnnnnnnnn nnnnnnnnnn ttcttggctt       60

tatatatctt gtggaaagga cgaaacaccg nnnnnnnnnn nnnnnnnnng ttttagagct      120

agaaatagca agttaaaata aggctagtcn nnnnnnnnnn nnnnnnnnnn nnnngtacaa      180

gcacacgttt gtcaagacc                                                   199


<210>  37
<211>  413
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  37
attcgccctt tgtacaaaaa agcaggcttt aaaggaacca attcagtcga ctggatccgg       60

taccaaggtc gggcaggaag agggcctatt tcccatgatt cttcatattt gcatatacga      120

tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagac      180

aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt      240

ttaaaatgga ctatcatatg cttaccgtaa cttgaagtat ttcgatttct tggctttata      300

tatcttaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt      360

gctttttttc tagacccagc tttcttgtac aaagttggca ttaaagggcg aat             413


<210>  38
<211>  70
<212>  DNA
<213>  Artificial

<220>
<223>  target oligonucleotide

<400>  38
taatacttta tctgtcccct ccaccccaca gtggggccac tagggacagg attggtgaca       60

gaaaagcccc                                                              70


