Gaetan Burgio
Deoxy Ribonucleic @GaetanBurgio Do you mean NgAgo?
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更新：研究者成功重复韩氏 NgAgo 方法
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呵呵，最新之后还有更新。韩氏 NgAgo 基因编辑法已经 通过PCR 验证在室温下成功用于编辑哺乳动物（老鼠）基因，研究者确认多处基因编辑-删除，并认为效率甚高（应该是与 CRISPR-Cas9 相比）。我不是内行，行话也是临时学，但字还是认识的，大家自己看啊。参见前文：《最新：研究者成功重复韩氏 NgAgo 方法》后我对事件的评论。下面是部分洋文的翻译。
【OK。讲座之后我在这发表了用 (韩氏）NgAgo 编辑老鼠的第一个 PCR 结果 --> 效率高！！！】（三个惊叹号系原文），
【你应该试试，非常高效（ PCR没有用 T7）】（You should try, this is the way to go. Highly efficient (the PCR was done without T7) 。
【正如那篇（韩春雨）的重大论文（seminal paper），（NgAgo）产生了多处基因编辑->删除】
@ren_chonghua Not really. I used one oligo. As per the seminal paper creates multiple editing => deletion
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仔细看看：确认能在室温工作。
#TAGC16 OK following the talk I put here my first PCR results on #CRISPR editing in mice using NgAg -> Efficient !!!
履历似乎比一篇 JBC （有质疑）的 方博高出1.5个量级
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@c0nc0rdance Sorry, yep!
@GaetanBurgio Haven't tried it yet. There's a small, forehead-shaped dent on my benchtop for current CRISPR project. #NHEJSucks
@c0nc0rdance You should try, this is the way to go. Highly efficient (the PCR was done without T7)
@GaetanBurgio Are you wanting to make deletions? Why there's some bands difference before T7E1 assay?
@ren_chonghua Not really. I used one oligo. As per the seminal paper creates multiple editing => deletion
@GaetanBurgio Do you mean you just used one gDNA but found some smaller bands by PCR?
@ren_chonghua Yes just used one p-oligo + NgAgo plasmid on a specific target. Amplified the genomic DNA by PCR. Control is lane 9. H2o is 10
This PCR results on NgAgo are prior to the T7 assay. Lane 1 to 7 sample Lane 8 C57BL/6 Lane 9 H2O
@GaetanBurgio What NgAgo clone did you use? The one from Addgene?
@scekker Correct from Addgene
@GaetanBurgio Do you use mutiple gDNA with NgAgo?
@zxtzhangqian Not for this experiment but am working on this
@GaetanBurgio Cool, I guess you tried NgAgo in mouse zygote, right? Look forward for more data from you:)
@zxtzhangqian Thanks! Yes these data are from mouse zygotes. More will hopefully come up soon.
@GaetanBurgio impressive will buy beer for details!
@BillPavan Ha Ha Ha will need quite a few :)
@GaetanBurgio can NgAgo work at temps lower than 37C?
@vectorgen I have no idea at all. We've just injected in zygotes at room To in M2 media
@GaetanBurgio room temp works, thanks!
@GaetanBurgio Congrats Gaetan! The community is having a heck of a time, would you share protocol info in this forum?https://groups.google.com/forum/#!topic/crispr/V1FALNbspCo …
@isaacbeing Yes happy to do that but I have to optimize my protocol first. I am afraid the mouse zygote is a ≠ beast to cell line
@GaetanBurgio Awesome, thank you! I know everyone is hungry for any info they can get
@GaetanBurgio you got deletions of a few hundred bp?
@YiquanTang No, probably ~50 to 100
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