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Journal article
Lentiviral Transduction-based CRISPR/Cas9 Editing of Schistosoma mansoni Acetylcholinesterase
Current Genomics, Vol.24(3), pp.155-170
2023
Abstract
Background: Recent studies on CRISPR/Cas9-mediated gene editing in Schistosoma mansoni have shed new light on the study and control of this parasitic helminth. However, the gene editing efficiency in this parasite is modest.
Methods: To improve the efficiency of CRISPR/Cas9 genome editing in schistosomes, we used lentivirus, which has been effectively used for gene editing in mammalian cells, to deliver plasmid DNA encoding Cas9 nuclease, a sgRNA targeting acetylcholinesterase (SmAChE) and a mCherry fluorescence marker into schistosomes.
Results: MCherry fluorescence was observed in transduced eggs, schistosomula, and adult worms, indicating that the CRISPR components had been delivered into these parasite stages by lentivirus. In addition, clearly changed phenotypes were observed in SmAChE-edited parasites, including decreased SmAChE activity, reduced hatching ability of edited eggs, and altered behavior of miracidia hatched from edited eggs. Next-generation sequencing analysis demonstrated that the lentiviral transduction-based CRISPR/Cas9 gene modifications in SmAChE-edited schistosomes were homology-directed repair predominant but with much lower efficiency than that obtained using electroporation (data previously published by our laboratory) for the delivery of CRISPR components.
Conclusion: Taken together, electroporation is more efficient than lentiviral transduction in the delivery of CRISPR/Cas9 into schistosomes for programmed genome editing. The exploration of tactics for enhancing CRISPR/Cas9 gene editing provides the basis for the future improvement of programmed genome editing in S. mansoni.
Details
- Title
- Lentiviral Transduction-based CRISPR/Cas9 Editing of Schistosoma mansoni Acetylcholinesterase
- Authors
- Xiaofeng Du (Author) - QIMR Berghofer Medical Research InstituteDonald P. McManus (Author) - The University of QueenslandJuliet D. French (Author) - QIMR Berghofer Medical Research InstituteHaran Sivakumaran (Author) - QIMR Berghofer Medical Research InstituteRebecca L. Johnston (Author) - QIMR Berghofer Medical Research InstituteOlga Kondrashova (Author) - QIMR Berghofer Medical Research InstituteConor E. Fogarty (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationMalcolm K. Jones (Author) - The University of QueenslandHong You (Corresponding Author) - QIMR Berghofer Medical Research Institute
- Publication details
- Current Genomics, Vol.24(3), pp.155-170
- Publisher
- Bentham Science Publishers Ltd.
- Date published
- 2023
- DOI
- 10.2174/1389202924666230823094608
- ISSN
- 1875-5488; 1389-2029
- Copyright note
- © 2023 The Author(s). Published by Bentham Science Publisher. This is an open access article published under CC BY 4.0 https://creativecommons.org/licenses/by/4.0/legalcode
- Data Availability
- The original contributions presented in the study are included in the article. Further inquiries can be directed to the corresponding author.
- Grant note
- DPM is a National Health and Medical Research Council (NHMRC) of Australia Leadership Fellow and receives Program (Grant no. APP1132975), Project (Grant no. APP1098244) and Investigator Grant (Grant no. APP1194462) support from the NHMRC for his research on schistosomes and schistosomiasis. XD holds a Research Training Program (RTP) Scholarship and Graduate School Scholarship from the University of Queensland, Australia. HY holds QIMR Berghofer Medical Research Institute Seed Funding Grants.
- Organisation Unit
- GeneCology Research Centre - Legacy; School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99982893702621
- Output Type
- Journal article
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