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

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)..

Department

Microbiology, Immunology, and Tropical Medicine

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