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Review of CRISPR/Cas9
- 1. CRISPR/Cas9 Thenewfrontierofgenome engineering 03.06.2015 JenniferA. Doudna & EmmanuelleCharpentier, Science346, 2014 DOI: 10.1126/science.1258096
- 7. 2phases 4
- 21. HowbacteriapreventDNAinvasionfromviruses CRISPR Protospacer 7Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 22. HowbacteriapreventDNAinvasionfromviruses CRISPR=ClusteredRegularly InterspacedShortPalindromic RepeatsCRISPR Protospacer Protospacer: InvadingDNAfrom viruses,phages,… 7Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 23. HowbacteriapreventDNAinvasionfromviruses CRISPR=ClusteredRegularly InterspacedShortPalindromic RepeatsCRISPR Protospacer Protospacer: InvadingDNAfrom viruses,phages,… 7Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 24. HowbacteriapreventDNAinvasionfromviruses tracRNA:transactivating CRISPRRNA tracRNA tracRNA 8Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 25. HowbacteriapreventDNAinvasionfromviruses tracRNA:transactivating CRISPRRNA tracRNA tracRNA 8Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 26. HowbacteriapreventDNAinvasionfromviruses tracRNA:transactivating CRISPRRNA tracRNA tracRNA 8Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 27. HowbacteriapreventDNAinvasionfromviruses 9Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 28. HowbacteriapreventDNAinvasionfromviruses 9Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 29. HowbacteriapreventDNAinvasionfromviruses tracRNAwill activatenuclease Cas9 9Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 30. HowbacteriapreventDNAinvasionfromviruses Cas9willsearchthe matchingforeignDNA tocreateDSBand promotedegradation tracRNAwill activatenuclease Cas9 9Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 31. HowbacteriapreventDNAinvasionfromviruses 9Sander et al, Nature Biotechnology, 32, 347-355, 2014 NewforeignDNAisaddedtoCRISPRregions
- 32. Cas9mechanism 10
- 34. Cas9mechanism PAM PAMmotif(‘NGG’) mandatorytocleaveDNA 2cleavagedomains: HNHandRuvC-like RuvC HNH 10
- 37. Asimplifiedsystem FusionofcrRNAandtracrRNAtoasinglegRNA (20 bp) 12Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 38. Asimplifiedsystem FusionofcrRNAandtracrRNAtoasinglegRNA (20 bp) 12Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 39. Asimplifiedsystem FusionofcrRNAandtracrRNAtoasinglegRNA 2-componentsystem (20 bp) 12Sander et al, Nature Biotechnology, 32, 347-355, 2014
- 57. TheRevolution
- 64. SomeApplications
- 66. DynamicImagingofgenomicloci Chen et al., Cell, 2013, Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system 21
- 67. DynamicImagingofgenomicloci Chen et al., Cell, 2013, Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system AttachedaGFPtoanuclease-deficient Cas9(dCas9) 21
- 68. DynamicImagingofgenomicloci Chen et al., Cell, 2013, Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system AttachedaGFPtoanuclease-deficient Cas9(dCas9) 21
- 69. DynamicImagingofgenomicloci Chen et al., Cell, 2013, Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system AttachedaGFPtoanuclease-deficient Cas9(dCas9) 21
- 71. Firstmonkeyswithcustomizedmutationsborn Niu et al., Cell, 2014, doi: 10.1016/j.cell.2014.01.027 22
- 72. Firstmonkeyswithcustomizedmutationsborn Niu et al., Cell, 2014, doi: 10.1016/j.cell.2014.01.027 22
- 73. Firstmonkeyswithcustomizedmutationsborn CRISPR/Cas9targeting ofmultiple genesinmonkeyembryos Ppar-gandRag1double mutationin monkeysinonestep Niu et al., Cell, 2014, doi: 10.1016/j.cell.2014.01.027 22
- 74. 23
- 76. Geneticallymodifyhumanembryos Liang et al., Protein Cell, 2015, CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes 23
- 77. Geneticallymodifyhumanembryos Liang et al., Protein Cell, 2015, CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Triedtomutatethehumanβ-globin(HBB)genein‘non-viable’ embryos(β-thalassaemia) 23
- 78. Geneticallymodifyhumanembryos Liang et al., Protein Cell, 2015, CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Triedtomutatethehumanβ-globin(HBB)genein‘non-viable’ embryos(β-thalassaemia) 7of86embryosweresuccessfullymutated Muchmorehigherratesofoff-targeting 23
- 79. Geneticallymodifyhumanembryos Liang et al., Protein Cell, 2015, CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Triedtomutatethehumanβ-globin(HBB)genein‘non-viable’ embryos(β-thalassaemia) 7of86embryosweresuccessfullymutated Muchmorehigherratesofoff-targeting Raisehugeethicalconcerns… 23
- 80. Conclusion
- 87. Thankyou