SummaryRMgm-1031
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Successful modification | The 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) : 24755823 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
Name parent line/clone | P. berghei ANKA cl15cy1 |
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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The mutant parasite was generated by | |
Name PI/Researcher | Manzoni G; Silvie O |
Name Group/Department | INSERM |
Name Institute | INSERM |
City | Paris |
Country | France |
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Name of the mutant parasite | |
RMgm number | RMgm-1031 |
Principal name | Δslarp-GFP-LUC |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not tested |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | intravital imaging of bioluminescent parasites was used to analyse the fate of Δslarp P. berghei sporozoites in vivo after intravenous inoculation into C57BL/6 mice. Control Δp230p-GFP-LUC parasites were readily detectable in the liver of infected mice 24 h after injection of sporozoites, and the signal further increased at 48 h. All mice developed a patent parasitemia, as shown by positive Giemsa-stained blood smears and diffusion of the luminescence signal in the entire mouse body at day 4 post-infection. In sharp contrast, mice injected with Δslarp-GFP-LUC showed no luminescence signal above background at any of the time points examined, and none of the animals became patent, in total agreement with published data showing that Δslarp P. berghei sporozoites fail to convert into blood stages. |
Additional remarks phenotype | Mutant/mutation Intravital imaging of bioluminescent parasites was used to analyse the fate of Δslarp P. berghei sporozoites in vivo after intravenous inoculation into C57BL/6 mice. Control Δp230p-GFP-LUC parasites were readily detectable in the liver of infected mice 24 h after injection of sporozoites, and the signal further increased at 48 h. All mice developed a patent parasitemia, as shown by positive Giemsa-stained blood smears and diffusion of the luminescence signal in the entire mouse body at day 4 post-infection. In sharp contrast, mice injected with Δslarp-GFP-LUC showed no luminescence signal above background at any of the time points examined, and none of the animals became patent, in total agreement with published data showing that Δslarp P. berghei sporozoites fail to convert into blood stages. Other mutants |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0902100 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1147000 | ||||||||||||||||||||||||
Gene product | sporozoite and liver stage asparagine-rich protein | sporozoite asparagine-rich protein | ||||||||||||||||||||||||
Gene product: Alternative name | SLARP; S22; SAP1 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | 5-fluorocytosine (5-FC) | ||||||||||||||||||||||||
Additional remarks genetic modification | The parasites have been generated and selected using the GOMO method of transfection (see RMgm-1026 for more details for this transfection method) | ||||||||||||||||||||||||
Additional remarks selection procedure | The parasites are selected by a combination of positive selection (pyrimethamine), negative selection (5-FC) and FACS sorting. 1) Transfected parasites are first selected in a mouse by pyrimethamine treatment 2) GFP+mCherry+ parasites are selected by FACS sorting and used to infect a mice 3) This mouse is treated with 5-FC to select for parasites that have the selectable marker removed 4) GFP+mCherry- and marker free parasites are selected by FAC sorting and used to infect a mouse | ||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: not Plasmodium | ||||||||||||||||||
Transgene name | GFP-Luciferase | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||
Type of plasmid/construct | Plasmid double cross-over | ||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||
Selection (negative) procedure | 5-fluorocytosine (5-FC) | ||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||
Additional remarks selection procedure | The parasites are selected by a combination of positive selection (pyrimethamine), negative selection (5-FC) and FACS sorting. 1) Transfected parasites are first selected in a mouse by pyrimethamine treatment 2) GFP+mCherry+ parasites are selected by FACS sorting and used to infect a mice 3) This mouse is treated with 5-FC to select for parasites that have the selectable marker removed 4) GFP+mCherry- and marker free parasites are selected by FAC sorting and used to infect a mouse | ||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PBANKA_1133400 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1357100 | ||||||||||||||||||
Gene product | elongation factor 1-alpha | ||||||||||||||||||
Gene product: Alternative name | EF-1alpha | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0719300 | ||||||||||||||||||
Gene product | bifunctional dihydrofolate reductase-thymidylate synthase, putative | ||||||||||||||||||
Gene product: Alternative name | dhfr/ts | ||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Replacement locus | ||||||||||||||||||
Gene Model of Parasite | PBANKA_0902100 | ||||||||||||||||||
Gene product | sporozoite and liver stage asparagine-rich protein sporozoite asparagine-rich protein | ||||||||||||||||||
Gene product: Alternative name | SLARP; S22; SAP1 | ||||||||||||||||||
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