EGFP reporter gene driven by ubiquitin promoter integration into Schistosoma mansoni gene safe harbor mediated by CRISPR/Cas
The genomic DNA was extracted from the multi-organism S. mansoni parasite after CRISPR/Cas gene editing on gene safe harbor (GSH) with 5' chemically modified-DNA donor and short homology arms. Briefly, schistosome parasites; mature worms or eggs were delivered with CRISPR component in the ribonucleoprotein complex (RNP) including cas9 or cas12a enzyme and and guide RNA targeting GSH site. The transgene donor delivered at the same time with RNP by electroporation. Then, several days post transfection, genomic DNAs were extracted and target amplicon were sequenced by next generation sequencing. The mutation resulting by CRISPR/Cas were estimated by RGEN tools and CRISPRressov2 softwares.