Summary

RMgm-1217
Malaria parasiteP. berghei
Genotype
Other modificationGene model (rodent): PBANKA_1445600; Gene model (P.falciparum): PF3D7_1230900; Gene product: serine/threonine protein kinase RIO1, putative (RIO1)
Details modification: Successful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants
Phenotype Asexual bloodstage;
Last modified: 21 March 2015, 21:26
  *RMgm-1217
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Other
Reference (PubMed-PMID number) Reference 1 (PMID number) : 25732065
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherGomes AR; Billker O
Name Group/DepartmentWellcome Trust Sanger Institute
Name InstituteWellcome Trust Sanger Institute
CityHinxton Cambridge
CountryUK
Name of the mutant parasite
RMgm numberRMgm-1217
Principal name-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageReduced fitness (growth rate)
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Successful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants.

Evidence is provided for reduced fitness (growth rate) of blood stages

In this study P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest.
In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants.
In this study parasites were (mainly) transfected with a pool of vectors targeting protein kinase genes for deletion.
Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis).

To analyze growth curves derived from barcode counting two parameters were considered: (1) the relative abundance of each barcode within the pool, and (2) the relative fitness of each mutant, i.e., the rate at which its abundance changed each day (i.e. growth rate).


  Other: Mutant parasite with another genetic modification
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1445600
Gene Model P. falciparum ortholog PF3D7_1230900
Gene productserine/threonine protein kinase RIO1, putative
Gene product: Alternative nameRIO1
Description
Short description of the modificationSuccessful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants
DescriptionIn this study P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest.
In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants.
In this study parasites were (mainly) transfected with a pool of vectors targeting protein kinase genes for deletion.
Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis).

The gene was targeted by PlasmoGEM vector PbGEM-065291