Design and Validation of CRISPR/Cas9 Systems for Targeted Gene Modification in Induced Pluripotent Stem Cells

Ciaran M Lee; Haibao Zhu; Timothy H Davis; Harshahardhan Deshmukh; Gang Bao

doi:10.1007/978-1-4939-6472-7_1

Design and Validation of CRISPR/Cas9 Systems for Targeted Gene Modification in Induced Pluripotent Stem Cells

Methods Mol Biol. 2017:1498:3-21. doi: 10.1007/978-1-4939-6472-7_1.

Authors

Ciaran M Lee¹, Haibao Zhu¹, Timothy H Davis¹, Harshahardhan Deshmukh¹, Gang Bao²

Affiliations

¹ Department of Bioengineering, Rice University, 6500 Main St., Houston, TX, 77030, USA.
² Department of Bioengineering, Rice University, 6500 Main St., Houston, TX, 77030, USA. gang.bao@rice.edu.

PMID: 27709565
DOI: 10.1007/978-1-4939-6472-7_1

Abstract

The CRISPR/Cas9 system is a powerful tool for precision genome editing. The ability to accurately modify genomic DNA in situ with single nucleotide precision opens up new possibilities for not only basic research but also biotechnology applications and clinical translation. In this chapter, we outline the procedures for design, screening, and validation of CRISPR/Cas9 systems for targeted modification of coding sequences in the human genome and how to perform genome editing in induced pluripotent stem cells with high efficiency and specificity.

Keywords: CRISPR; Genome editing; Targeted gene knockout.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics*
Cell Line
Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
Gene Editing / methods
Genome, Human / genetics*
HEK293 Cells
Humans
Pluripotent Stem Cells / physiology*
RNA Editing / genetics*

Grants and funding

PN2 EY018244/EY/NEI NIH HHS/United States