RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
MutatedGene model (rodent): PBANKA_0501200; Gene model (P.falciparum): PF3D7_1016900; Gene product: early transcribed membrane protein 10.3 | protein of early gametocyte 4 (UIS4, up-regulated in infective sporozoites, ETRAMP10.3)
Details mutation: Pbuis4 gene replaced with P. chabaudi uis4 gene (PCHAS_0501300)
PhenotypeNo phenotype has been described
Last modified: 8 November 2021, 17:28
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene mutation
Reference (PubMed-PMID number) Reference 1 (PMID number) : 34715088
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/ResearcherBrandsma AM, Montagna GN
Name Group/DepartmentParasitology Unit
Name InstituteMax Planck Institute for Infection Biology
Name of the mutant parasite
RMgm numberRMgm-5103
Principal nameuis4::Pcuis4
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

In the mutant the uis4 gene has been replaced by the uis4 gene of P. chabaudi (PCHAS_0501300). This gene is under control of the 3' and 5' UTR regions of uis4 of P. berghei. In addition, the mutant expresses GFP under control of the constitutive eef1a promoter

Protein (function)
UIS4  is expressed throughout liver stage development and localizes to the parasitophorous vacuole. UIS4 has a total length of 220 amino acid residues and can be divided into three domains: an amino-terminal domain of 54 residues including the signal peptide encompassing 25 residues, the trans-membrane region and a positively charged carboxy-terminal domain of 143 residues. Inside an infected hepatocyte, UIS4 is distributed at the PVM and the tubovesicular network (TVN), which forms vesicular protrusions into the host cell cytoplasm.

Phenotype in all life cycle stages not different from wild type P. berghei parasites 

Additional information
From the Abstract:
'Early transcribed membrane proteins (ETRAMPs) form a unique protein family in malaria parasites. These molecules are expressed during Plasmodium intracellular phases and inserted at the parasite parasitophorus vacuole membrane (PVM), which constitutes the host-parasite interface. Upregulated in infectious sporozoites 4 (UIS4) is an essential ETRAMP of liver stages of the murine malaria model parasite Plasmodium berghei.We generated transgenic P. berghei parasites where UIS4 was replaced by Plasmodium falciparum ETRAMP8 or ETRAMP10.3. Both ETRAMPs were expressed in transgenic parasite lines, but liver stage maturation was impaired, indicating that the selected ETRAMPs failed to substitute the function of UIS4. As a control, we included the UIS4 ortholog from the murine parasite Plasmodium chaubaudi. We observed that PcUIS4 successfully restores UIS4 function in P. berghei'

PfETRAMP8, PfETRAMP10.3 and PfETRAMP14.2 are the only members of the P. falciparum ETRAMP family expressed in sporozoites. In contrast to UIS4, PfETRAMP10.3 is also expressed in blood stages and is considerably shorter in amino acid length (Table 1). Previous work demonstrated that PfETRAMP10.3 did not complement the function of UIS4 in P. yoelii parasites. 

Other mutants

  Mutated: Mutant parasite with a mutated gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0501200
Gene Model P. falciparum ortholog PF3D7_1016900
Gene productearly transcribed membrane protein 10.3 | protein of early gametocyte 4
Gene product: Alternative nameUIS4, up-regulated in infective sporozoites, ETRAMP10.3
Details of the genetic modification
Short description of the mutationPbuis4 gene replaced with P. chabaudi uis4 gene (PCHAS_0501300)
Inducable system usedNo
Short description of the conditional mutagenesisNot available
Additional remarks inducable system
Type of plasmid/construct(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
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationFor generation of uis4::Pcuis4 transgenic parasites, the 5’ UTR of PbUIS4, the 3’ UTR of PbUIS3, and the 3’ UTR of PbUIS4 were amplified from P. berghei WT ANKA-GFP gDNA by PCR using the following primer combinations: UIS4-F2-EcoRI and UIS4-R2-SpeI, UIS4-F3-SphI and UIS4-R3-PvuII, and UIS4-F4-HindIII and UIS4-R4-KpnI, respectively. These fragments were digested with the indicated restriction enzymes and successively cloned into the PB386_pBAT1-G6 vector, resulting in the pBATint vector. Next, the ORF of PcUIS4 was amplified from Plasmodium chabaudi gDNA by PCR using the PcUIS4-F and PcUIS4-R primers, respectively. The ORF fragment was digested with SpeI and NaeI and cloned into the pBATint vector. The vector was subsequently linearized with SacI and ApaLI
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