Targeted insertion and reporter transgene activity at a gene safe harbor of the human blood fluke, Schistosoma mansoni

Authors

• Wannaporn Ittiprasert, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• Max F. Moescheid, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• Cristian Chaparro, IHPE, University of Perpignan Via Domitia, CNRS, IFREMER, University Montpellier, Perpignan, France.
• Victoria H. Mann, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• Thomas Quack, Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany.
• Rutchanee Rodpai, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• André Miller, Schistosomiasis Resource Center, Biomedical Research Institute, Rockville, MD 20850, USA.
• Prapakorn Wisitpongpun, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• Watunyoo Buakaew, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
• Margaret Mentink-Kane, Schistosomiasis Resource Center, Biomedical Research Institute, Rockville, MD 20850, USA.
• Sarah Schmid, Schistosomiasis Resource Center, Biomedical Research Institute, Rockville, MD 20850, USA.
• Anastas Popratiloff, Nanofabrication and Imaging Center, Science & Engineering Hall, George Washington University, Washington, DC 20052, USA.
• Christoph G. Grevelding, Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany.
• Christoph Grunau, IHPE, University of Perpignan Via Domitia, CNRS, IFREMER, University Montpellier, Perpignan, France.
• Paul J. Brindley, Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.

Document Type

Journal Article

Publication Date

7-24-2023

Journal

Cell reports methods

Volume

3

Issue

7

DOI

10.1016/j.crmeth.2023.100535

Keywords

Schistosoma mansoni; gene safe harbor; human blood fluke; overlapping CRISPR target; reporter transgene

Abstract

The identification and characterization of genomic safe harbor sites (GSHs) can facilitate consistent transgene activity with minimal disruption to the host cell genome. We combined computational genome annotation and chromatin structure analysis to predict the location of four GSHs in the human blood fluke, , a major infectious pathogen of the tropics. A transgene was introduced via CRISPR-Cas-assisted homology-directed repair into one of the GSHs in the egg of the parasite. Gene editing efficiencies of 24% and transgene-encoded fluorescence of 75% of gene-edited schistosome eggs were observed. The approach advances functional genomics for schistosomes by providing a tractable path for generating transgenics using homology-directed, repair-catalyzed transgene insertion. We also suggest that this work will serve as a roadmap for the development of similar approaches in helminths more broadly.

APA Citation

Ittiprasert, Wannaporn; Moescheid, Max F.; Chaparro, Cristian; Mann, Victoria H.; Quack, Thomas; Rodpai, Rutchanee; Miller, André; Wisitpongpun, Prapakorn; Buakaew, Watunyoo; Mentink-Kane, Margaret; Schmid, Sarah; Popratiloff, Anastas; Grevelding, Christoph G.; Grunau, Christoph; and Brindley, Paul J., "Targeted insertion and reporter transgene activity at a gene safe harbor of the human blood fluke, Schistosoma mansoni" (2023). GW Authored Works. Paper 3035.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/3035

Department

Microbiology, Immunology, and Tropical Medicine