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Direct production of mutants using ZFNs, TALENs or CRISPR/Cas in one-cell embryos
Published on Sep 04, 20141619 Views
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Chapter list
Direct production of mouse mutants using ZFNs, TALENs or CRISPR/Cas9 in one-cell embryos00:00
Mouse models New Mouse Models through Nuclease & Zygote Technology - made by gene targeting in ES cells since 1989 00:32
Single step mutagenesis by using nucleases in one-cell embryos02:31
Genomic editing: different strategies03:51
Roadmap04:20
Proof-of-principle using a zinc-finger nuclease in zygotes04:51
Rab38 ZFNs for the generation/correction of chocolate mutants06:51
ZFNs08:56
Method of the year 201109:41
What are TALs?09:57
The structure of TAL proteins10:48
The structure of TALENs - 111:12
The structure of TALENs - 211:37
The structure of TALENs - 312:26
TALEN construction kits13:54
TALEN construction - 114:56
TALEN construction - 215:40
IDG-TALENs - construction Kit17:05
TALEN Nuclease Activity - 118:24
TALEN Nuclease Activity - 219:33
Gene editing by TALENs in one-cell embryos20:28
Correction of the G19V Mutation in Cht21:30
C9orf72 Knockout Mice21:58
DHTKD1 Knockout23:34
Fus codon replacement mutants24:15
Optimizing gene editing events by TALENs in one-cell embryos25:57
Fus alleles identified in tail DNA are transmitted via the germline29:29
HRM Analysis of Fus- and Rik-TALENs off-target sites31:15
Systematic analysis of TALEN off-target activity33:20
Big genomic deletions using TALENs34:16
TALENs35:39
The CRISPR/Cas9 type II bacterial defense system36:18
What‘s CRISPR/Cas?37:25
The CRISPR/Cas9 system in Genome Engineering - 138:14
The CRISPR/Cas9 system in Genome Engineering - 239:41
Comparison of TALEN/Cas9 Nuclease activity40:43
Gene editing with Cas9 in Embryos - 141:28
Gene editing with Cas9 in Embryos - 242:05
CRISPR/Cas mediated editing of Rab38 in one-cell embryos42:26
Large genomic deletions: Transfected Neuro2A cells43:01
Targeting of two sites with sgRNAs to create genomic deletions44:00
Increasing CRISPR/Cas-specificity45:11
CRISPR/Cas-specificity analysis45:55
Types of Nickases47:25
Nickases activity48:24
Improvements to increase the number of targeted mice50:04
CRIPSR/Cas 51:08
ZFN,TALEN & CRIPSR/Cas mutagenesis51:32
The team51:57
Thanks for your attention52:08