Chromosome splitting of Plasmodium berghei using the CRISPR/Cas9 system

Daniel Addo-Gyan; Haruka Matsushita; Enya Sora; Tsubasa Nishi; Masao Yuda; Naoaki Shinzawa; Shiroh Iwanaga

doi:10.1371/journal.pone.0260176

Chromosome splitting of Plasmodium berghei using the CRISPR/Cas9 system

PLoS One. 2022 Feb 24;17(2):e0260176. doi: 10.1371/journal.pone.0260176. eCollection 2022.

Authors

Daniel Addo-Gyan¹, Haruka Matsushita¹, Enya Sora¹, Tsubasa Nishi², Masao Yuda², Naoaki Shinzawa¹, Shiroh Iwanaga^{3

4}

Affiliations

¹ Department of Environmental Parasitology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
² Laboratory of Medical Zoology, Department of Medicine, Mie University, Kurimamachi Yacho, Tsu, Mie, Japan.
³ Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Yamadaoka, Suita, Osaka, Japan.
⁴ Center for Infectious Disease Education and Research (CIDER), Osaka University, Yamadaoka, Suita, Osaka, Japan.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems / genetics*
Centromere / genetics
Chromosomes / genetics*
Gene Expression Regulation / genetics
Malaria / genetics*
Malaria / parasitology
Plasmodium berghei / genetics*
Plasmodium berghei / pathogenicity
Plasmodium falciparum / genetics
Plasmodium falciparum / pathogenicity
Rodentia / parasitology
Telomere / genetics

Grants and funding

All studies were supported by Grants-in-Aid for Scientific Research (20H03477 and 19K22527 to S.I. 18K07084 and 21K06985 to N.S., and 17H01542 to M.Y.), which were funded by the Japan Society for the Promotion of Science (JSPS: https://www.jsps.go.jp/), and also supported by the Japan Agency for Medical Research and Development (AMED: https://www.amed.go.jp/) under Grant Number 21jm0210061h0004 (to S.I.), 21wm0325018 (to N.S.) and 21wm0225014 (to N.S.). The funders have no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.