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DOI :
10.3791/55916-v
•
11:53 min
July 12th, 2017
Chapters
0:05
Title
0:50
Cloning of gRNAs into the CRISPER-concatemer Vector
3:40
Assessing Success of Cloning by Restriction Digest
5:28
Preparing Intestinal Organoids for Electroporation
8:08
Electroporation
10:36
Results: Successful Generation of CRISPER-concatemers and Electroporation of Intestinal Organoids
11:24
Conlcusion
このプロトコルは、CRISPR / Cas9技術を用いたマルチ遺伝子ノックアウトの作成に特に有用な、1つのガイドRNAコンカテマーベクターに複数の単一ガイドRNAをクローニングするための工程を記載する。腸オルガノイドにおける二重ノックアウトの生成は、この方法の適用可能性として示されている。
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