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Stem Cells International
Volume 2016, Article ID 2574152, 11 pages
Research Article

Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development

1Section of Hematology and Oncology, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA
2Center for Regenerative Medicine (CReM), Boston University and Boston Medical Center, Boston, MA 02118, USA
3Department of Environmental Health, Boston University School of Public Health, Boston, MA 02118, USA

Received 4 January 2016; Accepted 17 March 2016

Academic Editor: Fanny L. Casado

Copyright © 2016 Brenden W. Smith et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplemental Figure 1: Sanger sequencing confirms PCR validation strategy of CYP1A1 reporter iPSCs. (A) PCR products discussed in Figure 1C were purified and sequenced using the Sanger method (Genewiz, Inc.). Both amplicons include endogenous regions of the CYP1A1 locus as well as elements of the reporter construct (eGFP in the 5′ amplicon; puro resistance gene in the 3′ amplicon). (B) Full sequences for the Left Homology Arm, Right Homology Arm, and single guide RNA (sgRNA).

Supplemental Figure 2: CYP1A1 reporter iPSCs elicit a response to the AHR agonist FICZ. (A) CYP1A1 transcript level expression was increased in both the parental cell line (blue bar) and the targeted clone (red bar) in response to escalating doses of 6-formylindolo(3,2-b)carbazole (FICZ). (B) Luciferase expression was observed as a result of FICZ treatment exclusively in the targeted clone (red bar) but not in the parental cell line (blue bar), showing that the integrated reporter construct was faithfully reporting on AHR-dependent CYP1A1 upregulation.

  1. Supplementary Material