Unknown

Dataset Information

0

Highly efficient CRISPR/Cas9 system in Plasmodium falciparum using Cas9-expressing parasites and a linear donor template.


ABSTRACT: The CRISPR/Cas9 system is a powerful genetic engineering technology for Plasmodium falciparum. We here report further improvement of the CRISPR/Cas9 system by combining the Cas9-expressing parasite with a liner donor template DNA. The Cas9-expressing parasite was generated by inserting the cas9 gene in the genome by double crossover recombination. The site-directed mutagenesis and the fusion of fluorescence protein was achieved within two weeks with high efficiency (> 85%), by transfecting the schizonts of the Cas9-expressing parasite with the liner donor template and the plasmid carrying the sgRNAs. Notably, there were neither off-target mutations in the resultant transgenic parasites nor unexpected recombination, that are the technical problems of the current CRISPR/Cas9 system. Furthermore, with our system, two genes on different chromosomes were successfully modified in single transfection. Because of its high efficiency and robustness, our improved CRISPR/Cas9 system will become a standard technique for genetic engineering of P. falciparum, which dramatically advances future studies of this parasite.

SUBMITTER: Nishi T 

PROVIDER: S-EPMC8445982 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC7406498 | biostudies-literature
| S-EPMC4199390 | biostudies-literature
| S-EPMC9093057 | biostudies-literature
| S-EPMC6085629 | biostudies-literature
| S-EPMC6007333 | biostudies-other
| S-EPMC5324397 | biostudies-literature
2016-07-14 | GSE84342 | GEO
| S-EPMC6973022 | biostudies-literature
| S-EPMC7708629 | biostudies-literature