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Crispr-Cas9 |
allgemeines |
|
| Cas9 |
Bakterien-Enzym zur Abwehr von Phagen. Erkennt fremde DNA und zerschneidet sie. |
| CRISPR |
Clustered Regularly Interspaced Palindromic Repeats |
palindromisch: DNA-Abschnitte, deren Stränge gegenläufig die gleiche Sequenz aufweisen. |
| Cas |
CRISPR-associated Protein |
CAS fügt kleine DNA-Fragmente von Phagen zwischen
2 CRISPR in das Bakteriengenom ein. |
Diese Phasen-DNA wird in RNA übersetzt und dient CAS dazu, fremde DNA zu erkennen, um sie dann zu zerschneiden. |
| Cas9 |
Bakterien-Enzym zur Abwehr von Phagen. Erkennt fremde DNA und zerschneidet sie. |
| CRISPR-Cas |
CRISPR erkennt gehäuft auftretende, gleichmäßig verteilte Wiederholungen und CAS zerscheidet die RNA(1). Auch für Genmanipulation benutzt.
|
| 6 CRISPR/Cas-Typen |
19 Untertypen |
| Typ II |
CRISPR/Cas9 |
Spacer-Sequenzen werden in RNA-Moleküle umsetzt. |
Die RNA-Moleküle leiten eine Endonuklease zu der eindringenden Fremd-DNA hin, so dass es diese zerschneidet. |
| Typ VI |
Enzyme zerlegen die RNA statt DNA. |
Es fehlen die palindromischen Wiederholungen und der Apparat zum Einbau der Spacer. |
| Typ III |
Am häufigsten |
Reagieren auf den Prozess des Umschreibens von DNA in RNA |
| EcoRI |
Restriktionsenzyme |
Schneidet die DNA an der spezifischen Erkennungssequenz GAATTC.
| AA/CGTT AclI |
| A/AGCTT HindIII HindIII-HF® |
|
AAT/ATT SspI SspI-HF® |
| /AATT MluCI |
| A/CATGT PciI |
| A/CCGGT AgeI AgeI-HF® |
| ACCTGC(4/8) BspMI BfuAI |
| A/CCWGGT SexAI |
| A/CGCGT MluI MluI-HF® |
| ACGGC(12/14) BceAI |
| A/CGT HpyCH4IV |
| ACN/GT HpyCH4III |
| (10/15)ACNNNNGTAYC(12/7) BaeI |
| (9/12)ACNNNNNCTCC(10/7) BsaXI |
| A/CRYGT AflIII |
| A/CTAGT SpeI SpeI-HF® |
| ACTGG(1/-1) BsrI |
| ACTGGG(5/4) BmrI |
| A/GATCT BglII |
| AGC/GCT AfeI |
| AG/CT AluI |
| AGG/CCT StuI |
| AGT/ACT ScaI-HF® |
| AT/CGAT ClaI BspDI |
| ATCTATGTCGGGTGCGGAGAAAGAGGTAAT(-15/-19) PI-SceI |
| ATGCA/T NsiI NsiI-HF® |
| AT/TAAT AseI |
| ATTT/AAAT SwaI |
| (11/13)CAANNNNNGTGG(12/10) CspCI |
| C/AATTG MfeI MfeI-HF® |
| CACGAG Nb.BssSI |
| CACGAG(-5/-1) BssSI-v2 |
| CACGTC(-3/-3) BmgBI |
| CAC/GTG PmlI |
| CACNNN/GTG DraIII-HF® |
| CACNN/NNGTG AleI-v2 |
| CAGCAG(25/27) EcoP15I |
| CAG/CTG PvuII PvuII-HF® |
| CAGNNN/CTG AlwNI |
| CAGTG(2/0) BtsIMutI |
| CA/TATG NdeI |
| /CATG FatI |
| C/ATG CviAII |
| CATG/ NlaIII |
| CAYNN/NNRTG MslI |
| CC(12/16) FspEI |
| CCANNNNN/NNNNTGG XcmI |
| CCANNNNN/NTGG BstXI |
| CCANNNN/NTGG PflMI |
| CCATC(4/5) BccI |
| C/CATGG NcoI NcoI-HF® |
| CCCAGC(-5/-1) BseYI |
| CCCGC(4/6) FauI |
| C/CCGGG XmaI TspMI |
| CCC/GGG SmaI |
| (0/-1)CCD Nt.CviPII |
| CCDG(10/14) LpnPI |
| CCGC(-3/-1) AciI |
| CCGC/GG SacII |
| CCGCTC(-3/-3) BsrBI |
| C/CGG MspI HpaII |
| /CCNGG StyD4I |
| CC/NGG ScrFI |
| C/CNNGG BsaJI |
| CCNNNNN/NNGG BslI |
| C/CRYGG BtgI |
| CC/SGG NciI |
| C/CTAGG AvrII |
| CCTC(7/6) MnlI |
| CCTCAGC(-5/-7) Nt.BbvCI |
| CCTCAGC Nb.BbvCI |
| CCTCAGC(-5/-2) BbvCI |
| CCTGCA/GG SbfI SbfI-HF® |
| CCTNAGC(-5/-2) Bpu10I |
| CC/TNAGG Bsu36I |
| CCTNN/NNNAGG EcoNI |
| CCTTC(6/5) HpyAV |
| /CCWGG PspGI |
| CC/WGG BstNI |
| C/CWWGG StyI-HF® |
| (10/12)CGANNNNNNTGC(12/10) BcgI |
| CGAT/CG PvuI PvuI-HF® |
| CG/CG BstUI |
| C/GGCCG EagI EagI-HF® |
| CG/GWCCG RsrII |
| CGRY/CG BsiEI |
| C/GTACG BsiWI BsiWI-HF® |
| CGTCTC BsmBI-v2 |
| CGTCTC(1/5) Esp3I |
| CGWCG/ Hpy99I |
| CMG/CKG MspA1I |
| CNNNNNNNNNNN/NNNNNNNNNG AbaSI |
| CNNR(9/13) MspJI |
| CR/CCGGYG SgrAI |
| C/TAG BfaI |
| CTCAG(9/7) BspCNI |
| C/TCGAG XhoI PaeR7I |
| CTCTTC(1/4) EarI |
| CTGAAG(16/14) AcuI |
| CTGCA/G PstI PstI-HF® |
| CTGGAG(16/14) BpmI |
| C/TNAG DdeI |
| C/TRYAG SfcI |
| C/TTAAG AflII |
| CTTGAG(16/14) BpuEI |
| C/TYRAG SmlI |
| C/YCGRG AvaI BsoBI |
| GAAGA(8/7) MboII |
| GAAGAC(2/6) BbsI BbsI-HF® |
| GAANN/NNTTC XmnI |
| GAATGC(1/-1) BsmI |
| GAATGC Nb.BsmI |
| G/AATTC EcoRI EcoRI-HF® |
| GACGC(5/10) HgaI |
| GAC/GTC ZraI |
| GACGT/C AatII |
| GACN/NNGTC Tth111I PflFI |
| GACNN/NNGTC PshAI |
| GACNNN/NNGTC AhdI |
| GACNNNN/NNGTC DrdI |
| GAGCT/C SacI SacI-HF® |
| GAG/CTC Eco53kI |
| GAGGAG(10/8) BseRI |
| GAGTC(4/-5) Nt.BstNBI |
| GAGTC(5/5) MlyI |
| GAGTC(4/5) PleI |
| G/ANTC HinfI |
| GAT/ATC EcoRV EcoRV-HF® |
| GA/TC DpnI |
| /GATC MboI Sau3AI DpnII |
| GATNN/NNATC BsaBI |
| G/AWTC TfiI |
| GCAATG Nb.BsrDI |
| GCAATG(2/0) BsrDI |
| GCAGC(8/12) BbvI |
| GCAGTG Nb.BtsI |
| GCAGTG(2/0) BtsI-v2 |
| GCANNNN/NTGC BstAPI |
| GCATC(5/9) SfaNI |
| GCATG/C SphI SphI-HF® |
| GCCC/GGGC SrfI |
| GCCGAG(21/19) NmeAIII |
| G/CCGGC NgoMIV |
| GCC/GGC NaeI |
| GCCNNNN/NGGC BglI |
| GCGAT/CGC AsiSI |
| GCGATG(10/14) BtgZI |
| GCG/C HhaI |
| G/CGC HinP1I |
| G/CGCGC BssHII |
| GC/GGCCGC NotI NotI-HF® |
| GC/NGC Fnu4HI |
| GCN/NGC Cac8I |
| GCNNNNN/NNGC MwoI |
| GCTAG/C BmtI BmtI-HF® |
| G/CTAGC NheI NheI-HF® |
| GCTCTTC(1/-7) Nt.BspQI |
| GCTCTTC(1/4) SapI BspQI |
| GC/TNAGC BlpI |
| G/CWGC TseI ApeKI |
| GDGCH/C Bsp1286I |
| GGATC(4/-5) Nt.AlwI |
| GGATC(4/5) AlwI |
| G/GATCC BamHI BamHI-HF® |
| GGATG(9/13) FokI |
| GGATG(2/0) BtsCI |
| GG/CC HaeIII |
| GGCCGG/CC FseI |
| GGCCNNNN/NGGCC SfiI |
| G/GCGCC KasI |
| GGCGC/C PluTI |
| GG/CGCC NarI |
| GGC/GCC SfoI |
| GG/CGCGCC AscI |
| GGCGGA(11/9) EciI |
| GGGAC(10/14) BsmFI |
| GGGCC/C ApaI |
| G/GGCCC PspOMI |
| G/GNCC Sau96I |
| GGN/NCC NlaIV |
| G/GTACC Acc65I |
| GGTAC/C KpnI KpnI-HF® |
| GGTCTC(1/5) BsaI BsaI-HF®v2 |
| GGTGA(8/7) HphI |
| G/GTNACC BstEII BstEII-HF® |
| G/GWCC AvaII |
| G/GYRCC BanI |
| GKGCM/C BaeGI |
| GR/CGYC BsaHI |
| GRGCY/C BanII |
| GT/AC RsaI |
| G/TAC CviQI |
| GTATAC BstZ17I-HF® |
| GTATCC(6/5) BciVI |
| G/TCGAC SalI SalI-HF® |
| GTCTC(1/-5) Nt.BsmAI |
| GTCTC(1/5) BsmAI BcoDI |
| G/TGCAC ApaLI |
| GTGCAG(16/14) BsgI |
| GT/MKAC AccI |
| GTN/NAC Hpy166II |
| /GTSAC Tsp45I |
| GTT/AAC HpaI |
| GTTT/AAAC PmeI |
| GTY/RAC HincII |
| GWGCW/C BsiHKAI |
| NNCASTGNN/ TspRI |
| R/AATTY ApoI ApoI-HF |
| RCATG/Y NspI |
| R/CCGGY BsrFI-v2 |
| R/GATCY BstYI |
| RGCGC/Y HaeII |
| RG/CY CviKI-1 |
| RG/GNCCY EcoO109I |
| RG/GWCCY PpuMI |
| TAACTATAACGGTCCTAAGGTAGCGAA(-9/-13) I-CeuI |
| TAC/GTA SnaBI |
| TAGGGATAACAGGGTAAT(-9/-13) I-SceI |
| T/CATGA BspHI |
| T/CCGGA BspEI |
| TCCRAC(20/18) MmeI |
| T/CGA TaqI-v2 |
| TCG/CGA NruI NruI-HF® |
| TCN/GA Hpy188I |
| TC/NNGA Hpy188III |
| T/CTAGA XbaI |
| T/GATCA BclI BclI-HF |
| TG/CA HpyCH4V |
| TGC/GCA FspI |
| TGGCAAACAGCTATTATGGGTATTATGGGT(-13/-17) PI-PspI |
| TGG/CCA MscI |
| T/GTACA BsrGI BsrGI-HF® |
| T/TAA MseI |
| TTAAT/TAA PacI |
| TTA/TAA PsiI-v2 |
| TT/CGAA BstBI |
| TTT/AAA DraI |
| VC/TCGAGB PspXI |
| W/CCGGW BsaWI |
| YAC/GTR BsaAI |
| Y/GGCCR EaeI |
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| Quellen |
1.) Fonfara I, et al.:
Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems.
Nucleic Acids Research, doi: 10.1093/nar/gkt1074; 2013
2.) Nunez J K, et al.:
Integrase-Mediated Spacer Acquisition during CRISPR-Cas Adaptive Immunity.
Nature 2015;519:193–198
3.) Shipman S L, et al.:
Molecular Recordings by Directed CRISPR Spacer Acquisition.
Science 2016;353:aaf1175
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