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

Shenglan Li; Haipeng Xue; Bo Long; Li Sun; Tai Truong; Ying Liu

doi:10.3791/52539

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

JoVE Journal
Биология развития

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JoVE Journal Биология развития

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

10,993 Views

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14:46 min

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May 28, 2015

DOI:

10.3791/52539-v

DOI:

10.3791/52539-v

•

14:46 min

May 28, 2015

•

1 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

Резюме

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.