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CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models
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Capitoli
- 00:04Introduction
- 01:06Gene Targeting of Embryonic Stem Cells (ESCs)
- 04:16PCR Genotyping of Targeted ESCs
- 07:02Preparation of Eight-Cell Stage Embryo and Microinjection of ESCs
- 09:49Results: Screening of Targeted ESC Clones
- 10:10Conclusion
Here we present a protocol for developing genetically modified mouse models using embryonic stem cells, especially for large DNA knock-in (KI). This protocol is tuned up using CRISPR/Cas9 genome editing, resulting in significantly improved KI efficiency compared with the conventional homologous recombination-mediated linearized DNA targeting method.
