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通过CAS9蛋白-gRNA复合物电穿孔直接基因敲除Axolotl脊髓神经干细胞
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JoVE Journal
Genetics
Direct Gene Knock-out of Axolotl Spinal Cord Neural Stem Cells via Electroporation of CAS9 Protein-gRNA Complexes
Please note that all translations are automatically generated.
Click here for the English version.
通过CAS9蛋白-gRNA复合物电穿孔直接基因敲除Axolotl脊髓神经干细胞
DOI:
10.3791/59850-v
•
09:10 min
•
July 09, 2019
•
Wilson Pak-Kin Lou*
1,2
,
Liqun Wang*
3
,
Cheng Long
,
Lei Liu
,
Ji-Feng Fei
1
School of Life Sciences
,
South China Normal University
,
2
The Research Institute of Molecular Pathology (IMP)
,
Vienna Biocenter (VBC)
,
3
Institute for Brain Research and Rehabilitation (IBRR)
,
South China Normal University
Chapters
00:04
Title
01:16
CAS9-gRNA Ribonucleoprotein Mixture and Agarose Plate Preparation
02:33
Electroporator Configuration and Glass Microinjection Capillary Loading
04:09
Pneumatic Pump Configuration
04:37
CAS9-gRNA Ribonucleoprotein Mixture Injection
06:05
Electroporation
06:45
Knock-Out Efficiency Assessment
07:25
Results: Representative Knock-Out Effects in Axolotl Spinal Cord Neural Stem Cells
08:33
Conclusion
Summary
Automatic Translation
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Automatic Translation
这里介绍的一个协议,通过将CAS9-gRNA复合物注射到脊髓中央管中,然后进行电穿孔,在axolotl脊髓中执行时间和空间限制基因敲除。
Tags
Axolotl
Spinal Cord
Neural Stem Cells
Gene Knock-out
Electroporation
CAS9
GRNA
Regeneration
Molecular Mechanisms
Neural Stem Cell Function
Spinal Cord Injury
Agarose Electroporation Plate
Poring Pulse
Transfer Pulse
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