CRISPR/Cas9 Genome Editing of Human-Induced Pluripotent Stem Cells Followed by Granulocytic Differentiation

Benjamin Dannenmann; Masoud Nasri; Karl Welte; Julia Skokowa

doi:10.1007/978-1-0716-0290-4_27

CRISPR/Cas9 Genome Editing of Human-Induced Pluripotent Stem Cells Followed by Granulocytic Differentiation

Methods Mol Biol. 2020:2115:471-483. doi: 10.1007/978-1-0716-0290-4_27.

Authors

Benjamin Dannenmann¹, Masoud Nasri¹, Karl Welte¹, Julia Skokowa²

Affiliations

¹ Department of Oncology, Hematology, Immunology, and Rheumatology, University Hospital Tuebingen, Tuebingen, Germany.
² Department of Oncology, Hematology, Immunology, and Rheumatology, University Hospital Tuebingen, Tuebingen, Germany. Julia.Skokowa@med.uni-tuebingen.de.

PMID: 32006418
DOI: 10.1007/978-1-0716-0290-4_27

Abstract

Research on patient-derived induced pluripotent stem cells (iPSCs) could immensely benefit from the implementation of CRISPR/Cas9 genome editing of iPSCs, creating unique opportunities such as the establishment of isogenic iPSC lines for disease modeling or personalized patient-specific drug screenings. Here we describe a stepwise protocol of safe, efficient, and selection-free CRISPR/Cas9-mediated gene correction or knockout in human iPSCs followed by 3D spin-embryoid body (EB)-based hematopoietic/neutrophilic iPSC-differentiation.

Keywords: CRISPR/Cas9; Genome editing; Hematopoietic, and neutrophilic differentiation of iPSCs; Human-induced pluripotent stem cells; Ribonucleoprotein; iPSC differentiation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems*
Cell Line
Electroporation / methods
Gene Editing / methods*
Granulocytes / cytology*
Granulocytes / metabolism
Hematopoiesis*
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism
Transfection / methods