Multiplexed CRISPR-Cas9 protocol for large transgene integration into the Schistosoma mansoni genome

Authors

Wannaporn Ittiprasert, Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC 20037, USA. Electronic address: wannaporni@gwu.edu.
Max M. Moescheid, Institute of Parasitology, Biomedical Research Center Seltersberg (BFS), Justus Liebig University, 35392 Giessen, Germany.
Victoria H. Mann, Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC 20037, USA.
Paul J. Brindley, Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC 20037, USA. Electronic address: pbrindley@gwu.edu.

Document Type

Journal Article

Publication Date

2-13-2024

Journal

STAR protocols

Volume

5

Issue

1

DOI

10.1016/j.xpro.2024.102886

Keywords

CRISPR; Gene Expression; Model Organisms; Molecular Biology

Abstract

Precise, on-target CRISPR-Cas9 genome editing has been shown in Schistosoma mansoni, involving both non-homology end joining and homology-directed repair pathways. Here, we present a multiplexed CRISPR-Cas9 protocol for large transgene integration into the S. mansoni genome. We describe steps for deploying multiplexed ribonucleoprotein complexes (RNPs) and donor DNA preparation. We then detail procedures for introducing RNPs into schistosome eggs by square-wave electroporation in the presence of a 5' phosphorothioate-modified double-stranded donor transgene. For complete details on the use and execution of this protocol, please refer to Ittiprasert et al. (2023)..

APA Citation

Ittiprasert, Wannaporn; Moescheid, Max M.; Mann, Victoria H.; and Brindley, Paul J., "Multiplexed CRISPR-Cas9 protocol for large transgene integration into the Schistosoma mansoni genome" (2024). GW Authored Works. Paper 4350.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/4350

Department

Microbiology, Immunology, and Tropical Medicine