RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5359
Malaria parasiteP. chabaudi
Genotype
Transgene
 Transgene not Plasmodium: mCherry
Promoter: Gene model: PCHAS_1322800; Gene model (P.falciparum): PF3D7_1455800; Gene product: LCCL domain-containing protein (LAP4; LCCL/lectin adhesive-like protein 4; CCp2)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Replacement locus: Gene model: PCHAS_0308200; Gene product: 6-cysteine protein P230p, putative (P230p)
Phenotype Gametocyte/Gamete;
Last modified: 30 May 2023, 11:45
  *RMgm-5359
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 37076932
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. chabaudi
Parent strain/lineP. c.chabaudi AS
Name parent line/clone Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherCunningham DA, Langhorne J
Name Group/DepartmentThe Francis Crick Institute
Name InstituteThe Francis Crick Institute
CityLondon
CountryUK
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Name of the mutant parasite
RMgm numberRMgm-5359
Principal namePcAS(ccp2)mC230p pRBC (♀)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GametemCherry expressed in female gametocytes
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses mCherry under control of the female specific CCp2 (LCCL domain-containing protein) promoter

Protein (function)

Phenotype
mCherry expressed in female gametocytes

Additional information
This mutant has been used to separate/isolate female gametocytes for analysis/comparison of PIR gene expression in male and female gametocytes.

From the Abstract: 'Infected red blood cells containing female and male P. chabaudi gametocytes transcribe a distinct set of genes encoded by the multigene family pir. The overall patterns are similar to what has been observed in the close relative P. berghei, but here we show that gametocyte-associated pir genes are distinct from those involved in chronic blood-stage infection and highlight a male-associated pir gene'.

Other mutants

  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene namemCherry
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Details of the genetic modification
Inducable system usedNo
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 markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationFor the generation of specifically-tagged male and female gametocyte lines, promoters were chosen based on the published transcriptome for P berghei ANKA. A region of approximately 1 kb upstream of the translation start site of the dynein heavy chain (DHC) gene or of the CCp2 gene was selected, synthesized commercially (Genewiz), digested with AflII and BamH1 and used to replace the eef1a promoter to generate the plasmids PcDHC230pmC, and PcCCp2230pmC.
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PCHAS_1322800
Gene Model P. falciparum ortholog PF3D7_1455800
Gene productLCCL domain-containing protein
Gene product: Alternative nameLAP4; LCCL/lectin adhesive-like protein 4; CCp2
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PCHAS_0308200
Gene product6-cysteine protein P230p, putative
Gene product: Alternative nameP230p
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren