RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_0501400; Gene model (P.falciparum): PF3D7_1017100; Gene product: rhoptry neck protein 12 (RON12)
PhenotypeNo phenotype has been described
Last modified: 18 October 2018, 11:40
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 30290224
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/ResearcherOda-Yokouchi Y, Ishino T, Tsuboi T
Name Group/DepartmentDivision of Malaria Research, Proteo-Science Center
Name InstituteEhime University, Matsuyama
Name of the mutant parasite
RMgm numberRMgm-4544
Principal nameΔPbRON12
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

The mutant lacks expression of RON12

Protein (function)
Rhoptry neck protein 12 (RON12) is found only in Plasmodium and is highly conserved across the genus

No phenotype was detected throughout the complete life cycle that was different from wild type parasites

Additional information
Evidence is presented that RON12 is also expressed in sporozoites and that it is localized to the rhoptry body, rather than to the rhoptry neck as described for its localization pattern in merozoites.
RON12 remains at the apical end in sporozoites shortly after invasion of HepG2 cells, suggesting that RON12 is not secreted prior to invasion, which is like PfRON12 in merozoites. Within sporozoites transforming to the early liver stage, RON12 is localized in the cytoplasm as spots. At the late liver stage, punctate RON12 signal is additionally detected at the marginal region of parasites as well as the central area, but remain within the parasite rather than being secreted to the PV space. The signal intensity of RON12 increases during intra-hepatocytic development, suggesting that newly RON12 production occurs during liver stage parasite maturation.

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0501400
Gene Model P. falciparum ortholog PF3D7_1017100
Gene productrhoptry neck protein 12
Gene product: Alternative nameRON12
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationPbron12-disrupted parasites (ΔPbRON12) were generated by double crossover homologous recombination using the gene disruption vector pL0006 available from BEI Resources. To replace the endogenous pbron12 (PBANKA_0501400) genomic locus with a pyrimethamine-resistant selectable marker containing the human dihydrofolate reductase gene (dhfr), two homologous recombination cassettes (pbron12–5: −970 to +53 bp and pbron12–3: +466 to +1257) were inserted into pL0006 at both sides of the drug-resistant cassette. The DNA fragments of pbron12–5 (1023 bp) and pbron12–3 (792 bp) were amplified from PbANKA genomic DNA (gDNA) using PbRON12-KO5-F-HindIII (5′-CCCAAGCTTGTTGTTTGGATAATTGAGTTGCGT-3′) and PbRON12-KO5-R-BglII (5′-GAAGATCTACAACCAATATGCATACCAAAACC-3′), PbRON12-KO3-F-KpnI (5′-GGGGTACCGAAAATGGAGAAATATCTGAATCC-3′) and PbRON12-KO3-R-NheI (5′-CTAGCTAGCGACAACGTATATCTATACATATGA-3′), respectively. The fragments were inserted into HindIII and BglII sites or KpnI and NheI sites, respectively. The plasmid was digested with HindIII and NheI before transfection.
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6