Authors

G Rinaldi²; S SuttiprapaC CochranI J Tsai²; W IttiprasertV H MannN Holroyd²; S IordanskiyM I BukrinskyM Berriman ²; P J Brindley ¹ The George Washington University, United States;  ² Wellcome Trust Sanger Institute

Discussion

Functional genomic studies will facilitate the characterisation of newly genome sequences of schistosomes. VSVG-pseudotyped murine leukemia virus (MLV) can transduce eggs of Schistosoma mansoni leading to chromosomal integration and germline transmission of transgenes, facilitating the development of stable transgenic lines. Nonetheless, robust expression of transgenes has been a challenge; we are currently evaluating several strategies to overcome this issue. Lentiviruses, including VSVG-pseudotyped human immunodeficiency virus (HIV-1) likely can facilitate transgenesis of schistosomes. We showed that early steps of lentivirus infection including attachment of virions to the schistosome tegument, proviral cDNA synthesis, and genome integration take place. High throughput sequencing analyses revealed widespread genome integration of HIV. Transgenesis combined with the genome editing technology CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated 9) has shifted the landscape for manipulating the genome by introducing specific mutations in the DNA. New findings suggested that Cas9-gRNAs ribonucleoprotein complexes delivered into schistosomules by electroporation induced INDEL mutation in a model locus, the gene coding for interleukin-4 inducing principle of S. mansoni eggs (IPSE). Retroviral-based approaches coupled with CRISPR-Cas9-driven genome editing will facilitate functional genomics studies for this parasite.