DOI: 10.1101/2023.05.23.542002

Generation of a Plasmodium falciparum reporter line for studies of parasite biology throughout the life cycle

P.Suarez-Cortes G. Costa M. Andres D. Eyermann C. Kreschel L. Spohr C. Goosmann V. Brinkmann E. A. Levashina

P.Suarez-Cortes G. Costa M. Andres ...+5 E. A. Levashina

摘要

Fluorescence reporter strains of human malaria parasites are powerful tools to study the interaction of the parasites with both human and mosquito hosts. However, low fluorescence intensity in transmission-relevant parasite stages and the choice of insertion loci that cause parasite developmental defects in the mosquito largely limits usefulness of currently available lines. To overcome these limitations, we used a CRISPR-Cas9-mediated approach to generate PfOBC13GFP, a novel selection marker-free reporter parasite in the background of the African NF54 Plasmodium falciparum line. As docking site, we selected the OBC13 locus that is dispensable for asexual and sexual development in vitro. PfOBC13GFP parasites encode GFP flanked by hsp70 UTRs that drive strong fluorescence reporter expression throughout blood and mosquito stages, enabling parasite detection by such high throughput methods as flow cytometry. When compared to the parental line, PfOBC13GFP parasites showed normal development during blood and mosquito stages, and they efficiently infected the main African vector Anopheles coluzzii, overcoming one of the limitations of the previously developed fluorescent reporter lines based on the Pfs47 locus. PfOBC13GFP constitutes a potent tool enabling host-pathogen studies throughout Plasmodium life cycle.

展开全部

图表提取

论文十问

参考文献

被引用

提问与回答

暂无人提供速读十问回答

被引用

笔记

问答