| Title: |
A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei |
| Authors: |
Jonsdottir, Thorey K.; Paoletta, Martina; Ishizaki, Takahiro; Hernandez, Sophia Raine C.; Ivanova, Maria; Herrera Curbelo, Alicia; Saiki, Paulina A.; Selinger, Martin; Das, Debojyoti; Henriksson, Johan; Bushell, Ellen |
| Publisher Information: |
Umeå universitet, Molekylär Infektionsmedicin, Sverige (MIMS); Umeå universitet, Institutionen för molekylärbiologi (Medicinska fakulteten); Umeå universitet, Umeå Centre for Microbial Research (UCMR); Instituto de Agrobiotecnología y Biología Molecular (IABIMO), INTA–CONICET, de Los Reseros y Dr. Nicolás Repetto s/n, P.O. Box 25 (B1712WAA),Hurlingham, Buenos Aires, Argentina; Parasitology and Zoology Unit, Department of Infection and Pathology, School of Veterinary Medicine, Rakuno Gakuen University, 582 Bunkyodai-midorimachi, Hokkaido, Ebetsu, Japan; Division of Children’s and Women’s Health (BKH), Department of Biomedical and Clinical Sciences (BKV), Linköping University, Sjukhusvägen Building 511, Linköping, Sweden; IceLab, Umeå University, Umeå, Sweden |
| Publication Year: |
2025 |
| Collection: |
Umeå University: Publications (DiVA) |
| Subject Terms: |
Cell and Molecular Biology; Cell- och molekylärbiologi |
| Description: |
Many Plasmodium genes remain uncharacterized due to low genetic tractability. Previous large-scale knockout screens have only been able to target about half of the genome in the more genetically tractable rodent malaria parasite Plasmodium berghei. To overcome this limitation, we have developed a scalable CRISPR system called P. berghei high-throughput (PbHiT), which uses a single cloning step to generate targeting vectors with 100-bp homology arms physically linked to a guide RNA (gRNA) that effectively integrate into the target locus. We show that PbHiT coupled with gRNA sequencing robustly recapitulates known knockout mutant phenotypes in pooled transfections. Furthermore, we provide an online resource of knockout and tagging designs to target the entire P. berghei genome and scale-up vector production using a pooled ligation approach. This work presents for the first time a tool for high-throughput CRISPR screens in Plasmodium for studying the parasite’s biology at scale. |
| Document Type: |
article in journal/newspaper |
| File Description: |
application/pdf |
| Language: |
English |
| Relation: |
Nucleic Acids Research, 0305-1048, 2025, 53:2; PMID 39844455; ISI:001402022200002 |
| DOI: |
10.1093/nar/gkaf005 |
| Availability: |
http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-235698; https://doi.org/10.1093/nar/gkaf005 |
| Rights: |
info:eu-repo/semantics/openAccess |
| Accession Number: |
edsbas.135F939E |
| Database: |
BASE |