Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e

1.0K Views

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07:31 min

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February 17th, 2023

DOI :

10.3791/64977-v

February 17th, 2023

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Shaohui Zheng¹^,², Fang Liang³, Yu Zhang¹^,², Ji-Feng Fei⁴, Wei Qin¹^,², Yanmei Liu¹^,²

¹Key Laboratory of Brain, Cognition and Education Science, Ministry of Education, ²Institute for Brain Research and Rehabilitation, Guangdong Key Laboratory of Mental Health and Cognitive Science, South China Normal University, ³Institute of Modern Aquaculture Science and Engineering, Guangdong Provincial Engineering Technology Research Center for Environmentally Friendly Aquaculture, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Sciences, South China Normal University, ⁴Department of Pathology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences

Summary

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PAM Less Base Editing

Zebrafish Models

Human Genetic Disease

ZSpRY ABE8e

Adenine To Guanine Conversion

Single nucleotide Variants

Pathogenic Characteristics

Pneumatic Microinjector

MRNA gRNA Mixture

Microinjector Setup

Chapters in this video

0:04

Introduction

1:02

Preparation of Microinjection Capillaries and Modified RNA

2:23

RNA Microinjection of Zebrafish Embryos

4:36

Efficiency Analysis of EditR Base Edits

6:08

Results: TSR2 Diamond Blackfan Zebrafish Mutation

6:41

Conclusion

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