Gene editing protocols for human induced pluripotent stem cells (iPSCs)

Whether studying normal human cell biology or a disease state, primary human cell models are not always available and can be challenging to work within the lab. iPSCs bridge this issue as they are a renewable model for karyotypically normal cells, can be manipulated in their pluripotent state, and differentiated into biologically relevant cell types.

ipsc diagram

To harness the power of iPSCs, the ability to edit their genomes is necessary for functional genomic studies, disease modeling, and endogenous tagging. Understanding the need for protocols for these applications, we developed a handful of resources for those new to the world of editing iPSCs with CRISPR. Details on the application notes, and download links can be found below.

1. Knockout your gene of interest - We demonstrate the ability to modify the genome and create functional protein knockout models within a human iPSC line. (Download now)

2. Generate a SNP change - Research shows an increasing number of human diseases have a genetic component. To study the phenotypic effects of these mutations, co-delivery of a Cas9 nuclease, guide RNA, and a short double-strand DNA donor with homology to the expected cut site is utilized. The CRISPR machinery cuts the DNA and uses the Homology Directed Repair (HDR) pathway to repair the lesion with the donor, which incorporates the new sequence. Editing the cells in this manner enables the generation of a SNP or correcting the change to a wild-type sequence. (Download now)

3. Tag an endogenous protein with a fluorescent reporter – Introduction of large tags through HDR-mediated gene editing enables applications in the laboratory, such as imaging the localization of fluorescently tagged proteins of interest, and generating reporter cell lines. (Download now)


Ryan Donnelly headshotWritten by Ryan Donnelly

He holds a Professional Science Masters in Molecular Biotechnology from George Washington University and Bachelors in Chemistry from the University of Florida.



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