RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1107
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1427200; Gene model (P.falciparum): PF3D7_0812300; Gene product: sporozoite surface protein 3, putative (SSP3)
Transgene
 Transgene not Plasmodium: GFPmut2
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
3'UTR: Gene model: Not available; Gene product: Not available
Replacement locus: Gene model: PY17X_1427200; Gene product: sporozoite surface protein 3, putative (SSP3)
Phenotype Sporozoite; Liver stage;
Last modified: 29 August 2014, 19:15
  *RMgm-1107
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 25156733
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone cl1.1
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherHarupa A; Kappe SHI
Name Group/DepartmentSeattle Biomedical Research Institute
Name InstituteSeattle Biomedical Research Institute
CitySeattle
CountryUSA
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Name of the mutant parasite
RMgm numberRMgm-1107
Principal namessp3(-)
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/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNormal numbers of sporozoites are formed. Gliding motility is affected. Sporozoites showed normal hepatocyte traversal and invasion in vitro. Hepatocyte invasion and liver stage development in vivo was comparable to wild type parasites.
Liver stageNormal numbers of sporozoites are formed. Gliding motility is affected. Sporozoites showed normal hepatocyte traversal and invasion in vitro. Hepatocyte invasion and liver stage development in vivo was comparable to wild type parasites.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of sporozoite surface protein 3, putative (SSP3) and expresses GFP under control of the constitutive eef1a promoter.

Protein (function)
SSP3 is a predicted type 1 transmembrane protein. BLASTP identified orthologs in all Plasmodium species, but did not yield any potential orthologs in other apicomplexan or non-apicomplexan organisms. SSP3 contains thirteen highly  conserved cysteine residues, twelve of which are within the predicted ectodomain and  one within the predicted transmembrane domain. Structural homology searches suggested that the N-terminal region of SSP3 is similar to the s48/45 domain of Plasmodium 6-Cys proteins.

Phenotype
Normal numbers of sporozoites are formed. Gliding motility is affected. Sporozoites showed normal hepatocyte traversal and invasion in vitro. Hepatocyte invasion and liver stage development in vivo was comparable to wild type parasites.

Additional information
Evidence is presented for a surface localisation on sporozoites (but is not shed in gliding trails during gliding motility).
Evidence is provided that SSP3 might be masked by CSP and is exposed and accessible to antibodies once CSP is shed.

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_1427200
Gene Model P. falciparum ortholog PF3D7_0812300
Gene productsporozoite surface protein 3, putative
Gene product: Alternative nameSSP3
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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 modificationNo information (sequence) of primers used to amplify the 5' and 3' targetting regions is provided
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
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  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFPmut2
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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 parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationNo information (sequence) of primers used to amplify the 5' and 3' targetting regions is provided
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_1133300
Gene Model P. falciparum ortholog PF3D7_1357100
Gene productelongation factor 1-alpha
Gene product: Alternative nameeef1a
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 Not available
Gene productNot available
Gene product: Alternative name
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 PY17X_1427200
Gene productsporozoite surface protein 3, putative
Gene product: Alternative nameSSP3
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