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

DOI :

10.3791/52539-v

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May 28th, 2015

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10,885 Views

Shenglan Li^1,2*, Haipeng Xue^1,2*, Bo Long^1,2,5, Li Sun^1,2,6, Tai Truong^1,2,4,7, Ying Liu^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

Chapters

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.

Tags

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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