Chromosome splitting of Plasmodium berghei using the CRISPR/Cas9 system

Spatial arrangement of chromosomes is responsible for gene expression in Plasmodium parasites. However, methods for rearranging chromosomes have not been established, which makes it difficult to investigate its role in detail. Here, we report a method for splitting chromosome in rodent malaria parasite by CRISPR/Cas9 system using fragments in which a telomere and a centromere were incorporated. The resultant split chromosomes segregated accurately into daughter parasites by the centromere. In addition, elongation of de novo telomeres were observed, indicating its proper function. Furthermore, chromosome splitting had no effect on development of parasites. Splitting of the chromosome is expected to alter its spatial arrangement, and our method will thus be useful for investigating its biological role related with gene expression.

Automated summary

This study presents a method for splitting chromosomes in rodent malaria parasites using the CRISPR/Cas9 system. The split chromosomes were accurately segregated into daughter parasites and the elongation of de novo telomeres was observed, indicating their proper function. Chromosome splitting had no effect on parasite development, suggesting its potential as a tool for investigating the biological role of chromosome spatial arrangement in gene expression.