CRISPR/Cas9 mediated CXCL4 knockout in human iPS cells of polycythemia vera patient with JAK2 V617F mutation

Janik Boehnke; Salim Atakhanov; Marcelo A S Toledo; Herdit M Schüler; Stephanie Sontag; Nicolas Chatain; Steffen Koschmieder; Tim H Brümmendorf; Rafael Kramann; Martin Zenke

doi:10.1016/j.scr.2021.102490

CRISPR/Cas9 mediated CXCL4 knockout in human iPS cells of polycythemia vera patient with JAK2 V617F mutation

Stem Cell Res. 2021 Aug:55:102490. doi: 10.1016/j.scr.2021.102490. Epub 2021 Aug 5.

Authors

Affiliations

¹ Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Medical School, Aachen, Germany; Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
² Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Medical School, Aachen, Germany; Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany; Department of Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, RWTH Aachen University Hospital, Aachen, Germany; Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf (CIO ABCD), Aachen, Germany.
³ Institute for Human Genetics, RWTH Aachen University Hospital, Aachen, Germany.
⁴ Department of Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, RWTH Aachen University Hospital, Aachen, Germany; Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf (CIO ABCD), Aachen, Germany.
⁵ Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Medical School, Aachen, Germany.
⁶ Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Medical School, Aachen, Germany; Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany. Electronic address: martin.zenke@rwth-aachen.de.

PMID: 34391098
DOI: 10.1016/j.scr.2021.102490

Abstract

The chemokine CXCL4/platelet factor 4 (PF4) gene, a key player in myelofibrosis, was knocked out by CRISPR/Cas9 in induced pluripotent stem cells (iPS cells) of a polycythemia vera (PV) patient with JAK2 V617F mutation. Two CXCL4^KO iPS cell lines with and without JAK2 V617F mutation (UKAi002-B-1 and UKAi002-A-1, respectively) were generated. CXCL4^KO iPS cells showed deletion of exon 1 and complete loss of CXCL4 protein. Pluripotency of iPS cells was confirmed by expression of pluripotency markers and trilineage differentiation. CXCL4^KO iPS cells are expected to provide a valuable tool for investigating the role of CXCL4 in human diseases.

Keywords: CXCL4; Hematopoiesis; Induced pluripotent stem cell; JAK2 V617F; PF4; iPS cells.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics
Humans
Induced Pluripotent Stem Cells* / metabolism
Janus Kinase 2 / genetics
Janus Kinase 2 / metabolism
Mutation
Polycythemia Vera* / genetics

Substances

JAK2 protein, human
Janus Kinase 2