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Journal / Developmental Biology / Efficient Generation of hiPSC Neural Lineage…
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Developmental Biology

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JoVE Journal Developmental Biology
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Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

Article DOI: 10.3791/52539-v • 14:46 min • May 28th, 2015
May 28th, 2015 •
*1,2, *1,2, 1,2,5, 1,2,6, 1,2,4,7, 1,2,3
1Department of Neurosurgery, The University of Texas Health Science Center at Houston, 2Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 3The Senator Lloyd & B. A. Bentsen Center for Stroke Research, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 4Summer Research Program, Office of Educational Programs, The University of Texas Health Science Center at Houston, 5Department of Anesthesiology, Shengjing Hospital, China Medical University, 6Department of Oncology, Renji Hospital, Shanghai Jiaotong University School of Medicine, 7Biology Department, University of West Georgia
* These authors contributed equally

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Genome editing tools such as the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas (CRISPR-associated) system have greatly improved gene targeting efficiency in human induced pluripotent stem cells (hiPSCs). This manuscript describes a protocol for generating lineage specific hiPSC reporter using CRISPR/Cas system assisted homologous recombination.

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Gene Targeting CRISPR/Cas9 Cas9 Double Nickase System Genetic Manipulation Genome Editing Tools Human Induced Pluripotent Stem Cell (hiPSC) Reporters Homologous Recombination Targeting Vectors Single Guide RNAs Mutation Correction
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