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E16 랫 태아의 Utero의 심실 주입 및 Electroporation에

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Institute for Regeneration Medicine, University of California, San Francisco - UCSF

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Cite this Article: E16 랫 태아의 Utero의 심실 주입 및 Electroporation에

Walantus, W., Elias, L., Kriegstein, A. In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos. J. Vis. Exp. (6), e236, doi:10.3791/236 (2007).

Abstract: E16 랫 태아의 Utero의 심실 주입 및 Electroporation에

- utero에서 배아 쥐의 네크로 텍스의 인 - 생체내 주입과 electroporation은 측면 뇌실의 벽면 라이닝 개인 progenitors의 조작을위한 강력한 도구를 제공합니다. 이 기술은 현재 널리 이상 - 표현이나 유전자를 쓰러뜨린 및 세포 증식, 이주 및 분화에 미치는 영향을 관찰하여 corticogenesis에 관련된 프로세스를 연구하는 데 사용됩니다. 전통적인 녹아웃 전략에 비해에서 - utero의 electroporation은 특정 시간적 창 동안 세포의 인구를 조작하는 빠른 방법을 제공합니다. 이 비디오 프로토콜에서는, 우리는 수술에 대한 쥐 준비 laporatomy 통해 자궁 뿔이 노출, 개발 배아의 측면 심실로 DNA를 주입, 측면 벽을 라이닝 progenitors로 DNA를 electroporating하고, 동물을 돌보는을위한 실험 방법을 설명 후 수술. 우리 연구실이 동영상과 같이 그러나 그것이 가장 일반적으로 E16에서 수행, E15 - E21 쥐에서 수술이 프로토콜을 사용합니다.

Disclosures: E16 랫 태아의 Utero의 심실 주입 및 Electroporation에

Ask the Author: E16 랫 태아의 Utero의 심실 주입 및 Electroporation에

7 Comments

Very nice work. I do this routinely in Mouse but have little luck with Rat. I get poor survival of pups. If I do a sham experiment, I get fine survival with electroporation only but little survival if I inject only. I've used a variety of anesthetics. Any suggestions particular to Rat? Thank you much

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Posted by: AnonymousOctober 24, 2007, 2:50 PM

Try different methods of purifying the DNA.

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Posted by: AnonymousOctober 29, 2007, 5:19 PM

I am having poor survival as well. Any ideas about different methods of purification? I am currently using a Promega MIDI/Maxi kit. Have you noticed with purification methods are better?

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Posted by: Chris CunninghamOctober 18, 2010, 1:53 PM

hi barry. are you beveling your injection pipettes enough? if they are not sharp enough embryos will have high mortality. have a look at the fourth chapter of the video again.

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Posted by: Aaron K.October 31, 2007, 2:12 AM

Thanks, will try, but mice go just fine without problems except that B6 mice don't make the best of moms.

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Posted by: AnonymousOctober 31, 2007, 8:53 AM

Hi, I would appreciate if you could send me the protocol for mice ...Thanks a lot !
Martine
m.barkats@institut-myologie.org

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Posted by: Martine B.May 28, 2010, 11:40 AM

Where can we find the tools used to make the syrine injector tip?

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Posted by: DerekJuly 17, 2008, 2:59 PM

what are the medicine for motor neuron disease?

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Posted by: saraSeptember 8, 2009, 1:23 AM

From which company can I buy the electrods for the electroporation. The same as you have. Could you tell me their cat. # also?

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Posted by: Julie CardinNovember 10, 2009, 4:19 PM


BTX Harvard Apparatus Tweezertrodes, you can get them from Fisher and other vendors. -William

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Posted by: AnonymousNovember 10, 2009, 4:40 PM

I was not able to find the volume for these intraventricular infusions. Also, what is the maximum infusion volume for these kinds of procedures using newborn rats? Thanks!

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Posted by: mark b.August 7, 2010, 8:49 AM

I was not able to find the volume for these intraventricular infusions. Also, what is the maximum infusion volume for these kinds of procedures using newborn rats? Thanks!

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Posted by: mark b.August 7, 2010, 8:50 AM

Have you ever delivered DNA earlier into the lateral ventricles (E12.5 or E13)?

Thank you.

Shelley

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Posted by: Shelley RussekNovember 30, 2011, 11:22 PM

No. Sorry, There are some who do though. You can use ultrasound or IR videomicroscopy.

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Posted by: BarryDecember 1, 2011, 4:17 PM

Thank you...any one in particular that you know has a technique working or a paper published? Thanks!

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Posted by: ShelleyDecember 1, 2011, 4:56 PM

Thank you...any one in particular that you know has a technique working or a paper published? Thanks!

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Posted by: ShelleyDecember 1, 2011, 4:57 PM

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