Effect of two mismatches as function of position and separation on Cas9 cleavage activity The tables below contain the raw data plotted in bubble heatmaps where: max Δ abundance defined the gradient colour and abundance-min defined the bubble size for the supercoiled pool Δ abundance defined the gradient colour and abundance-max defined the bubble size for the nicked pool > Sp = SpCas9 Sa = SaCas9 HF1 = SpCas9 HF1 Hypa = HypaCas9 HiFi or IDT = IDT Alt-R HiFi Cas9 > 002 = pLibrary PS4 = KMlib002 003 = pLibrary EMX1 = KMlib003 > MM = mismatch > dist = distance between the two mismatches ************************************************** pLibrary PS4 - supercoiled pool max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the whole target length dist 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Sp max Δ abundance -0.55457665 -0.487685864 -0.527837795 -0.553603729 -0.620638739 -0.611501274 -0.642704757 -0.65321639 -0.642329634 -0.630760876 -0.6188233 -0.639069399 -0.660284803 -0.607991312 -0.637945181 -0.642189078 -0.606309552 -0.560005063 -0.665054699 abundance min 0.182146206 0.326374503 0.295509285 0.238735628 0.190212374 0.086755911 0.083920137 0.070816164 0.071593883 0.067307472 0.067632352 0.074632833 0.075411901 0.083323112 0.084453774 0.092862734 0.095162065 0.121013883 0.148164892 Sa max Δ abundance -0.397374655 -0.325209487 -0.372986011 -0.337350618 -0.354545118 -0.379672779 -0.3801009 -0.376717225 -0.386678615 -0.435374912 -0.432115666 -0.391632063 -0.410049365 -0.405248087 -0.368203846 -0.363538326 -0.246559377 -0.357303012 -0.37571806 abundance min 0.073618931 0.080606825 0.095701469 0.088000733 0.082338546 0.074124153 0.068862829 0.049001047 0.066400069 0.077806056 0.07576988 0.081247634 0.114087158 0.08525624 0.076949007 0.087064329 0.117306241 0.05249784 0.023593865 HF1 max Δ abundance -0.126593692 -0.142948619 -0.189875943 -0.140009752 -0.228748453 -0.196588808 -0.187096106 -0.150330742 -0.161221576 -0.176855665 -0.145950967 -0.149247585 -0.139729151 -0.131426885 -0.076514837 -0.093186936 -0.040607447 -0.042746084 -0.081833699 abundance min 0.202784143 0.288714349 0.258046497 0.220003284 0.22922151 0.189124877 0.16641733 0.170172843 0.164745507 0.192078931 0.187181648 0.207318542 0.248338371 0.290545885 0.341008523 0.309030845 0.332768571 0.303874289 0.173413823 Hypa max Δ abundance -0.191551525 -0.156769146 -0.199808752 -0.151497082 -0.1978001 -0.226675006 -0.261176186 -0.215640422 -0.204427802 -0.227435284 -0.228587771 -0.229782526 -0.212320685 -0.247809929 -0.169404494 -0.154960517 -0.137200122 -0.197071552 -0.247417276 abundance min 0.175001377 0.303068609 0.292131239 0.249955781 0.220475197 0.133988568 0.128253014 0.137649433 0.146982879 0.119347612 0.113847687 0.120323477 0.112677999 0.100829083 0.093678048 0.091659311 0.120121278 0.085274743 0.076139733 HiFi max Δ abundance -0.386968392 -0.320183664 -0.362804229 -0.392803134 -0.449355812 -0.457283204 -0.484161776 -0.480629366 -0.472424747 -0.474937992 -0.46729826 -0.473070322 -0.431977852 -0.417147035 -0.3575813 -0.334313164 -0.230788394 -0.252406885 -0.359634491 abundance min 0.189022287 0.342155388 0.308759861 0.238633402 0.21855426 0.091867929 0.083100631 0.058859556 0.057372877 0.065327106 0.06690585 0.06536773 0.060561493 0.07066946 0.069472157 0.07893858 0.074431639 0.095756596 0.083595691 max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the seed region dist 0 1 2 3 4 5 6 7 8 Sp max Δ abundance -0.45280068 -0.323625536 -0.420420674 -0.421974333 -0.488031444 -0.471907545 -0.555957026 -0.558664536 -0.374879264 abundance min 0.294464612 0.60466687 0.542790667 0.44974325 0.389676096 0.159621409 0.157900416 0.161825425 0.175774968 Sa max Δ abundance -0.384542697 -0.325470354 -0.37887555 -0.329343962 -0.338341274 -0.281272254 -0.243103898 -0.203908792 -0.338222921 abundance min 0.093757122 0.090771997 0.103227555 0.075567789 0.108407428 0.135214748 0.128046174 0.042163401 0.026837565 HF1 max Δ abundance -0.082169114 -0.104003116 -0.16356528 -0.075600277 -0.158899594 -0.096275007 -0.160630128 -0.08553989 -0.136635497 abundance min 0.230954694 0.406940478 0.326534872 0.24815912 0.306577925 0.220742694 0.125166542 0.146540978 0.092499037 Hypa max Δ abundance -0.171093601 -0.126538551 -0.176724412 -0.125451444 -0.159003613 -0.198551535 -0.255281005 -0.185347674 -0.151609467 abundance min 0.216958697 0.479675195 0.433876969 0.340501402 0.351951994 0.104553825 0.096973295 0.07756973 0.07724202 HiFi max Δ abundance -0.38655019 -0.226975687 -0.292077299 -0.285823884 -0.360989409 -0.334376765 -0.465138284 -0.416872249 -0.333517002 abundance min 0.29774107 0.627232749 0.562638178 0.45394803 0.426418147 0.131833872 0.118729898 0.074160957 0.117928442 max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the PAM-distal region dist 0 1 2 3 4 5 6 7 8 Sp max Δ abundance -0.657486992 -0.622257207 -0.615268137 -0.665826937 -0.701266697 -0.677632452 -0.727917208 -0.771122966 -0.785483998 abundance min 0.058354976 0.059460597 0.056438109 0.062047036 0.053013552 0.04911335 0.055810863 0.042254485 0.056808587 Sa max Δ abundance -0.384269353 -0.292115677 -0.341900558 -0.328916472 -0.356660126 -0.445471116 -0.481682948 -0.500459686 -0.527021044 abundance min 0.053795653 0.078222453 0.111318709 0.130863403 0.092147577 0.059112156 0.05239881 0.03486468 0.023923625 HF1 max Δ abundance -0.148803916 -0.170299051 -0.224267958 -0.169797229 -0.218174634 -0.202111166 -0.1534837 -0.096647978 -0.103230936 abundance min 0.17522731 0.187614515 0.211231205 0.233447956 0.241120091 0.265439672 0.330848569 0.295118003 0.132998628 Hypa max Δ abundance -0.202375077 -0.181173598 -0.225369033 -0.175897418 -0.182368582 -0.209470234 -0.196621552 -0.176488233 -0.234181051 abundance min 0.13220244 0.151205747 0.173737761 0.194443736 0.181664586 0.208171935 0.195911218 0.222149255 0.125503025 HiFi max Δ abundance -0.350031853 -0.361014604 -0.38918388 -0.393891387 -0.401105664 -0.392197344 -0.327435015 -0.250265266 -0.409352057 abundance min 0.070441327 0.079718972 0.075756435 0.07803499 0.095659947 0.102670756 0.119946676 0.082412402 0.055166075 ************************************************** pLibrary EMX1 - supercoiled pool max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the whole target length dist 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Sp max Δ abundance -0.633118025 -0.575837999 -0.572391265 -0.546646608 -0.601856764 -0.62707753 -0.677502386 -0.685986748 -0.702677676 -0.694857544 -0.69734641 -0.696581026 -0.691816698 -0.700839567 -0.668595802 -0.600079657 -0.616691102 -0.563311619 -0.590738097 abundance min 0.166325187 0.283339436 0.265353938 0.259542745 0.202644569 0.171552175 0.089896942 0.047312103 0.028850303 0.019703055 0.019025576 0.017797562 0.020196307 0.018186685 0.019888717 0.02354765 0.01990351 0.026598565 0.011395883 Sa max Δ abundance -0.297411751 -0.283731915 -0.279006644 -0.216041467 -0.208817232 -0.233567157 -0.281237446 -0.247362685 -0.218899635 -0.272390832 -0.229809495 -0.312364259 -0.273418855 -0.234141733 -0.219048849 -0.164903202 -0.263993415 -0.272899059 -0.16354435 abundance min 0.316165709 0.417049128 0.394579837 0.417775337 0.335295974 0.32110212 0.245253692 0.202659531 0.184434926 0.175849718 0.166939574 0.160213933 0.149943738 0.187978022 0.165506255 0.186436929 0.150037159 0.21036702 0.1277475 HF1 max Δ abundance -0.496179116 -0.479330755 -0.440380496 -0.416333705 -0.478692249 -0.535660484 -0.594338549 -0.586066829 -0.591093325 -0.599686002 -0.613796192 -0.638017092 -0.65363696 -0.672345991 -0.65922726 -0.668747686 -0.666267726 -0.676514753 -0.74615104 abundance min 0.136697881 0.262391217 0.214760707 0.244713919 0.168439991 0.147919868 0.079352756 0.064056856 0.058748268 0.048148671 0.051003258 0.056424201 0.046862436 0.051796459 0.047688316 0.052902298 0.036858963 0.045372513 0.030652924 Hypa max Δ abundance -0.453388302 -0.393893731 -0.400204819 -0.359370813 -0.422242256 -0.497418542 -0.557212794 -0.558760471 -0.595447658 -0.628595725 -0.64714064 -0.638435614 -0.675829241 -0.721396637 -0.784088249 -0.768628042 -0.795573932 -0.799313245 -0.875456169 abundance min 0.085691145 0.203530141 0.178588612 0.180790377 0.103375403 0.093957191 0.042568315 0.030540897 0.027299532 0.033897972 0.018494037 0.027422349 0.024503174 0.029632911 0.025086449 0.014950133 0.022106176 0.01825371 0.033521648 HiFi max Δ abundance -0.573786568 -0.517946954 -0.51568701 -0.493141054 -0.553615094 -0.581756238 -0.624499367 -0.624828974 -0.639659363 -0.649643039 -0.656024718 -0.650625247 -0.643022372 -0.670646197 -0.605653714 -0.563541626 -0.570784604 -0.491106719 -0.575674611 abundance min 0.126623969 0.245595414 0.211609329 0.230206843 0.16139787 0.133124751 0.067539167 0.039564977 0.029226266 0.026095798 0.020731148 0.026845494 0.020118736 0.019990605 0.025571255 0.022341219 0.024179244 0.015267299 0.019554032 max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the seed region dist 0 1 2 3 4 5 6 7 8 Sp max Δ abundance -0.407258431 -0.300586516 -0.286639122 -0.221583769 -0.268970099 -0.148452141 -0.264346841 -0.293384659 -0.480241171 abundance min 0.342677008 0.621501608 0.619935863 0.616229301 0.610466505 0.680692383 0.381136075 0.190657265 0.017921411 Sa max Δ abundance -0.253287842 -0.272665763 -0.265666066 -0.225662893 -0.136516416 -0.146219631 -0.207067938 -0.116415073 -0.359310541 abundance min 0.419590569 0.627573742 0.609007511 0.625757214 0.573186425 0.654780614 0.457851131 0.393324997 0.124693265 HF1 max Δ abundance -0.466577585 -0.405195421 -0.367508202 -0.341660419 -0.372098337 -0.334016431 -0.409353113 -0.566230625 -0.708369415 abundance min 0.167614264 0.459833155 0.382934119 0.452307205 0.33759161 0.382591433 0.154785586 0.046007333 0.013773243 Hypa max Δ abundance -0.423841997 -0.290323872 -0.272417237 -0.230860202 -0.249318911 -0.21007785 -0.369442968 -0.465131516 -0.764988399 abundance min 0.12018151 0.376427373 0.338251378 0.375310873 0.252036284 0.294020129 0.087889255 0.015839795 0.019722894 HiFi max Δ abundance -0.351547184 -0.264640121 -0.233316098 -0.197806114 -0.27193591 -0.137926208 -0.241851529 -0.273533864 -0.399341979 abundance min 0.252759755 0.528357845 0.476070927 0.53558748 0.473963894 0.517696549 0.275352937 0.128397968 0.023182838 max Δ abundance and abundance-min for target sequences with two misamtches separated by different number of nucleotides across the PAM-distal region dist 0 1 2 3 4 5 6 7 8 Sp max Δ abundance -0.809526269 -0.791362807 -0.80127782 -0.78461375 -0.79865894 -0.837535225 -0.843224601 -0.842579833 -0.794065823 abundance min 0.02801955 0.027032347 0.027750775 0.032091062 0.024781576 0.022203879 0.028845463 0.025295806 0.016395257 Sa max Δ abundance -0.318853588 -0.315949474 -0.326451963 -0.204633272 -0.278857245 -0.262424778 -0.321012951 -0.234016807 -0.122769796 abundance min 0.241832228 0.241069009 0.265984173 0.294689742 0.269860586 0.268932131 0.259608739 0.223736759 0.291868801 HF1 max Δ abundance -0.506220773 -0.488049631 -0.455340854 -0.415333126 -0.510245052 -0.586098274 -0.651995124 -0.615614202 -0.399854135 abundance min 0.121911852 0.126611214 0.113374027 0.137427228 0.133678743 0.113728979 0.07154852 0.069795083 0.079502657 Hypa max Δ abundance -0.460841292 -0.423812357 -0.437755671 -0.419472875 -0.549680338 -0.624717004 -0.674437301 -0.784092619 -0.634293121 abundance min 0.062712518 0.085888195 0.084577409 0.085641066 0.055443232 0.049017623 0.052201148 0.03338176 0.046999375 HiFi max Δ abundance -0.753554893 -0.731948877 -0.751193897 -0.718983868 -0.752132275 -0.774234139 -0.775705234 -0.777601088 -0.70367465 abundance min 0.028592732 0.030727519 0.034354209 0.035722886 0.022542931 0.022886496 0.022058494 0.012193435 0.013023189 ************************************************** ************************************************** ************************************************** pLibrary PS4 - nicked pool Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the whole target length dist 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Sp Δ abundance -0.014039728 -0.009468576 -0.009474021 -0.009080311 -0.024212535 -0.019998091 -0.016254671 -0.011683716 -0.000245145 -0.00268386 -0.000791046 0.001092921 0.003325953 -0.002791373 0.000852456 0.000348019 -0.000407708 0.005398112 -0.017795859 abundance max 0.179955527 0.168278102 0.164448289 0.167557371 0.166538708 0.165602019 0.159126199 0.138657954 0.125391362 0.131769013 0.128698491 0.128678793 0.126776237 0.126154105 0.148471163 0.140473168 0.117787899 0.122171441 0.163683882 Sa Δ abundance -0.066246512 -0.024364838 -0.043399298 -0.05692083 -0.021315678 -0.029788222 -0.059048773 -0.074069631 -0.097043002 -0.100121797 -0.057183197 -0.043043135 -0.043209237 -0.00485284 0.03490627 0.01765873 0.032556387 -0.099057363 -0.148115417 abundance max 0.423274787 0.389745208 0.416807958 0.446987985 0.40063162 0.437545617 0.47979616 0.518744279 0.500500288 0.462414888 0.361375677 0.331397377 0.294229665 0.329433427 0.460410664 0.336180809 0.407924269 0.475534013 0.518151337 HF1 Δ abundance 0.124452298 0.109483616 0.085781792 0.095828236 0.091425453 0.090022041 0.10983644 0.10509931 0.076987939 0.096033449 0.115237301 0.113032397 0.068347884 0.085110269 0.105823795 0.07873832 0.051576376 0.052476302 0.016178484 abundance max 0.47477005 0.462117278 0.486751461 0.538403335 0.506263188 0.565871784 0.570423155 0.507569955 0.488051577 0.509455731 0.527024373 0.492096979 0.423610772 0.444727186 0.583158518 0.430531498 0.416950595 0.391632651 0.432769536 Hypa Δ abundance 0.139848792 0.130515939 0.126120629 0.123578671 0.122382653 0.138253111 0.135774776 0.116394605 0.107091878 0.106297497 0.118797572 0.124920801 0.104130336 0.102742052 0.147774634 0.134809704 0.117393286 0.075040091 0.094805701 abundance max 0.392224896 0.354863871 0.359509625 0.388735742 0.394650622 0.424512116 0.39360739 0.370953739 0.342951982 0.33237399 0.3327131 0.371543434 0.321130859 0.320907459 0.436911673 0.363004285 0.323422543 0.259350693 0.316428359 HiFi Δ abundance 0.001968454 0.026505452 0.019099462 0.029701413 0.005699789 -0.000368833 -0.001772258 0.013467911 0.003885145 0.020499401 0.022114038 0.022873763 0.034101026 0.044388935 0.075172087 0.058481086 0.064573405 0.027857828 0.008335831 abundance max 0.362209932 0.319823221 0.300165283 0.325628471 0.30136931 0.310818138 0.295121882 0.307918496 0.274071947 0.298693579 0.300373988 0.308047925 0.303917846 0.344053398 0.380696467 0.336174976 0.315044619 0.250968844 0.246479836 Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the seed region dist 0 1 2 3 4 5 6 7 8 Sp Δ abundance 0.016402305 0.009136182 0.003038424 0.002203104 0.001863612 0.00408968 0.005439436 -0.003874228 -0.003432907 abundance max 0.176559845 0.208821543 0.197289504 0.19565953 0.171149866 0.1461455 0.143552351 0.121598312 0.213249273 Sa Δ abundance -0.079777151 -0.061072129 -0.109891733 -0.136977881 -0.056832808 -0.03243141 -0.008715652 0.010208444 -0.256516041 abundance max 0.4547892 0.394438927 0.487127813 0.571412908 0.43222369 0.396620839 0.426137371 0.748171105 1 HF1 Δ abundance 0.084339849 0.05344097 0.025128169 0.050006847 0.029675916 0.002592729 0.065620634 0.034006871 0.019519908 abundance max 0.406110496 0.336628634 0.332603058 0.349584833 0.342343577 0.360913826 0.436403476 0.4122161 0.489469362 Hypa Δ abundance 0.117245191 0.068208656 0.061047783 0.075792858 0.087279727 0.086449642 0.099839839 0.070192743 0.075227022 abundance max 0.353092055 0.258448386 0.272741761 0.301861192 0.330864933 0.334084749 0.339233233 0.340780376 0.337217075 HiFi Δ abundance 0.018798425 0.008724029 0.005188182 0.006087563 0.011426343 0.015053876 0.006564299 -0.000422888 -0.009027302 abundance max 0.328074686 0.203871925 0.187940071 0.214378506 0.243356972 0.300237121 0.268043184 0.215176851 0.25283334 Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the PAM-distal region dist 0 1 2 3 4 5 6 7 8 Sp Δ abundance -0.051965892 -0.019131511 -0.010501405 -0.004250076 -0.035472973 -0.016253775 -0.015626963 -0.034971736 -0.016853365 abundance max 0.225879116 0.208865508 0.190662774 0.191323368 0.175965468 0.150233751 0.143673688 0.149211673 0.112841406 Sa Δ abundance -0.030975274 0.028383063 0.030735154 0.022072116 0.053919812 0.013433627 -0.081561485 -0.198186187 -0.285374638 abundance max 0.330661905 0.335701964 0.326324179 0.360253532 0.430838254 0.465279598 0.542810585 0.779022786 0.907628955 HF1 Δ abundance 0.195747065 0.168707397 0.153753017 0.141273103 0.130121079 0.067549018 0.065198492 0.056781185 0.052118885 abundance max 0.648569633 0.6558517 0.685542015 0.685473976 0.627574706 0.487306658 0.402789961 0.424161662 0.351147242 Hypa Δ abundance 0.174558921 0.17466853 0.18553567 0.147044373 0.145042297 0.116179994 0.100307874 0.094424502 0.073735893 abundance max 0.443089603 0.415954457 0.44583274 0.426003934 0.497805302 0.372402949 0.359235234 0.306570215 0.298649842 HiFi Δ abundance -0.017637646 0.066908644 0.055556044 0.082960768 0.04233332 0.053327911 0.041107797 0.071483737 0.002706736 abundance max 0.415436039 0.455865316 0.408618318 0.442340539 0.369085762 0.327217943 0.362201446 0.327771912 0.291521999 ************************************************** pLibrary EMX1 - nicked pool Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the whole target length dist 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Sp Δ abundance -0.024210934 -0.018142335 -0.023827592 -0.021479228 -0.027016538 -0.027951805 -0.025264299 -0.030247408 -0.029648654 -0.029964666 -0.026087046 -0.029113829 -0.030531668 -0.033464827 -0.033377885 -0.024189002 -0.027409323 -0.022540778 -0.020672844 abundance max 0.13559149 0.141996156 0.151977115 0.135864488 0.145846166 0.149828737 0.138173239 0.143540169 0.145820623 0.147951951 0.153637417 0.153302555 0.15272573 0.15205396 0.154695987 0.147091437 0.156583683 0.145071806 0.140444911 Sa Δ abundance 0.000112148 0.010024199 0.006635194 -0.000389278 0.009322971 0.000494987 0.00856421 0.008393843 0.006167148 0.001377595 0.003968107 -0.007436144 0.000992678 -0.004650353 -0.002751774 0.01015735 -0.005490784 -0.003624031 0.003336878 abundance max 0.214461211 0.222421494 0.217143744 0.19561941 0.226608126 0.211440391 0.212995028 0.225508805 0.221144436 0.200144901 0.201569256 0.190243647 0.197796986 0.19227045 0.188326759 0.187240362 0.185939569 0.183769405 0.208967053 HF1 Δ abundance -0.000130159 0.010019275 0.004016257 -0.000930496 0.014275543 -0.013867945 -0.021261 -0.015095376 -0.018247036 -0.02215534 -0.021202757 -0.020015511 -0.028934582 -0.03606742 -0.041203492 -0.033645263 -0.041332117 -0.064278092 -0.051619303 abundance max 0.274405501 0.271464021 0.280507452 0.251115827 0.298065226 0.273255877 0.278094887 0.2591362 0.263876891 0.258162162 0.280631421 0.271197284 0.280566192 0.284220977 0.295746841 0.276662188 0.23557302 0.247152472 0.212372614 Hypa Δ abundance -0.015981618 -0.009939715 -0.013457782 -0.014143217 -0.011991235 -0.019164246 -0.025825487 -0.023953761 -0.030311613 -0.024426467 -0.029643247 -0.032021873 -0.037554353 -0.031572743 -0.028687406 -0.038838475 -0.029484281 -0.035459115 -0.041841727 abundance max 0.12582648 0.131629193 0.133188775 0.114049249 0.123112243 0.128639501 0.133704389 0.128524911 0.13240677 0.126215339 0.137021869 0.13788108 0.149898128 0.131287595 0.128069492 0.13068271 0.10787665 0.105110136 0.094779253 HiFi Δ abundance -0.004081706 0.001992174 0.002826537 -0.001614589 -0.007593909 -0.013721032 -0.008080635 -0.012292652 -0.009101067 -0.003176838 -0.004986117 0.001159874 -0.008929427 -0.007235121 -0.009257798 -0.006631494 0.002979967 0.008272365 0.004272565 abundance max 0.207704849 0.23596713 0.240422335 0.202534538 0.211700674 0.199871419 0.183764409 0.190170167 0.18595819 0.188862412 0.19442143 0.203727626 0.204436155 0.178324797 0.17055108 0.163015838 0.18206852 0.169359433 0.18516364 Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the seed region dist 0 1 2 3 4 5 6 7 8 Sp Δ abundance -0.013036143 -0.002198206 -0.001545844 -0.003931735 -0.002797667 -0.000965975 -0.002827202 -0.003448378 -0.028145439 abundance max 0.175296513 0.226608758 0.238652096 0.207543206 0.238257385 0.231456513 0.177420419 0.183219654 0.171651199 Sa Δ abundance 0.008418604 0.004296366 0.006647393 0.008344456 -0.010256167 0.000551322 0.013421567 0.030304395 -0.005906394 abundance max 0.205799447 0.194454173 0.217941091 0.189932526 0.198002279 0.217535159 0.197139776 0.250776029 0.224191277 HF1 Δ abundance -0.008150346 -0.005361806 -0.010434874 -0.017195978 0.010143348 -0.017619092 -0.023491737 -0.032391425 -0.050395821 abundance max 0.240967803 0.219721893 0.23526747 0.214693146 0.256933695 0.229681956 0.221977066 0.256966758 0.266378317 Hypa Δ abundance -0.021259844 -0.011628761 -0.012078814 -0.009518663 -0.015466952 -0.012474818 -0.014459055 -0.020114857 -0.025740848 abundance max 0.131661415 0.122111694 0.126990016 0.112479468 0.124508664 0.122477604 0.115952337 0.127417783 0.133967127 HiFi Δ abundance 0.033673925 0.031374797 0.040504433 0.026062703 0.037446507 0.032127663 0.026033182 0.029971305 0.013615982 abundance max 0.319467868 0.356474722 0.398349055 0.317735838 0.374286371 0.346609246 0.334515473 0.306441231 0.287616275 Δ abundance and abundance-max for target sequences with two misamtches separated by different number of nucleotides across the PAM-distal region dist 0 1 2 3 4 5 6 7 8 Sp Δ abundance -0.033572282 -0.031918818 -0.039015577 -0.035838451 -0.035588279 -0.041032651 -0.03427352 -0.040072631 -0.03847934 abundance max 0.113920729 0.122579925 0.126084644 0.116935375 0.101217399 0.107797238 0.088548908 0.102748037 0.10301636 Sa Δ abundance -0.008221678 -0.000769428 7.97726E-05 -0.011795997 -0.000646697 -0.0096151 -0.007706758 0.006300343 0.012769374 abundance max 0.21183481 0.216962202 0.216419332 0.219757341 0.213835794 0.237137928 0.245071987 0.253466685 0.283466018 HF1 Δ abundance 0.013224304 0.036342737 0.03695816 0.035277066 0.045742923 0.020318886 -0.010737241 -0.017441381 -0.033601752 abundance max 0.304698301 0.329987831 0.357347695 0.343701768 0.31703793 0.312789561 0.282548162 0.23026102 0.279036568 Hypa Δ abundance -0.009513955 -0.003397419 -0.007164947 -0.008772368 -0.001961023 -0.012789327 -0.024650741 -0.033761072 -0.046884119 abundance max 0.120169526 0.148236665 0.15082258 0.12942482 0.123955894 0.112201213 0.118779776 0.101708576 0.118657341 HiFi Δ abundance -0.037277282 -0.028945166 -0.037455383 -0.023575106 -0.036788365 -0.046664739 -0.040447085 -0.048837818 -0.039946484 abundance max 0.153903416 0.161906917 0.160521214 0.147900231 0.13513926 0.142722791 0.123231653 0.142365326 0.131985751