RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_1224500; Gene model (P.falciparum): PF3D7_0806200; Gene product: Dpy-19-like C-mannosyltransferase (CmanT, DPY19)
Phenotype Fertilization and ookinete; Oocyst; Sporozoite;
Last modified: 1 August 2022, 11:25
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) : 35594144
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/ResearcherSrivastava PN, Mishra S
Name Group/DepartmentDivision of Molecular Microbiology and Immunology
Name InstituteCSIR-Central Drug Research Institute
Name of the mutant parasite
RMgm numberRMgm-5209
Principal nameCmanT-
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 ookineteNormal, wild-type gametocyte and ookinete production. However, ookinetes showed strongly reduced motility and motion velocity. No oocysts are formed
OocystNo oocyst formation
SporozoiteNo oocyst and sporozoite formation
Liver stageNot tested
Additional remarks phenotype

The mutant lacks expression of CmanT and expresses GFP under control of the constitutive hsp70 promoter (the GFP reporter cassette is introduced into the disrupted CmanT locus)
Protein (function)
From the paper: 'C-Mannosylation of the thrombospondin type I repeat (TSR) domains is one of the most important factors involved in their function. It occurs on the first tryptophan of the WXXWXXC conserved motif where the tryptophan is usually surrounded by arginine or lysine forming the ligand-binding stretch of this sticky domain. It is found in its canonical or modified forms in many Plasmodium proteins. The Plasmodium proteome contains nine known canonical TSR domain-containing proteins. TSR containing proteins such as thrombospondin-like anonymous protein (TRAP), circumsporozoite protein (CSP), CSP and TRAP related protein (CTRP), and secreted protein with altered thrombospondin repeat (SPATR) have all been shown to be important for various parasite processes and life cycle stages. Here, we show that C-mannosylation catalyzing enzyme C-mannosyltransferase (CmanT) plays an essential role in malaria transmission in Plasmodium berghei.'


Normal, wild-type-like growth/multiplication of blood stages. Normal, wild-type-like gametocyte and ookinete production. However, ookinetes showed strongly reduced motility and motion velocity. No oocyst and sporozoite formation

Additional information
From the paper: 'CTRP was the first protein shown to be essential for ookinete motility and infectivity of mosquito midgut. Since the potential C-mannosylation motif is present in the TSR domain of CTRP and the phenotype of mutants lacking CTRP expression and the CmanT− mutants is similar, we hypothesize that C-mannosylation of the TSR domain of CTRP is one of the essential factors responsible for the CTRP mutants phenotype.'

See also the paper 'Tryptophan C-mannosylation is critical for Plasmodium falciparum transmission for mutants lacking DPY19' (PMID: 35906227). From the Abstract: 'DPY19-deficiency abolishes C-glycosylation, destabilizes members of the TRAP adhesin family and inhibits transmission to mosquitoes'. Mutants show defects in gametocyte egress, exflagellation, ookinete infectivity ond oocyst production (and mosquito infectivity). Evidence is presented that C-glycosylation is dispensable in P. falciparum salivary gland sporozoites (because O-fucosylation of essential TSR proteins like TRAP is sufficient to compensate for the absence of tryptophan C-mannosylation).

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1224500
Gene Model P. falciparum ortholog PF3D7_0806200
Gene productDpy-19-like C-mannosyltransferase
Gene product: Alternative nameCmanT, DPY19
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/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationTo replace PbCmanT with a targeting construct, two homology regions (F1, 5′ UTR and F2, 3′ UTR) were amplified using primers 1292/1293 and 1294/1295 and cloned at SalI and NotI/AscI, respectively, in the pBC-GFP-hDHFR:yFCU vector. The resulting construct was linearized with XhoI/AscI.
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