SummaryRMgm-76
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Successful modification | The 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) : 12471098 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei NK65 |
Name parent line/clone | RMgm-9 |
Other information parent line | The parent line used for the genetic modification is a mutant (RMgm-9) lacking the wild type CS gene |
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The mutant parasite was generated by | |
Name PI/Researcher | C. Persson, V. Nussenzweig, E. Nardin |
Name Group/Department | Departmentof Medical and Molecular Parasitology |
Name Institute | New York University School of Medicine |
City | New York |
Country | USA |
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Name of the mutant parasite | |
RMgm number | RMgm-76 |
Principal name | CS(pf) |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Normal numbers of of mature oocysts in A. stephensi mosquitoes. Salivary gland infectivity and the number of sporozoites per salivary gland was reduced compared to wild type (~30% of wild type). |
Sporozoite | Salivary gland infectivity and the number of sporozoites per salivary gland was reduced compared to wild type (~30% of wild type). Mutant sporozoites display normal morphology and motility. The sporozoites were found to invade and develop within HepG2 cells comparable to wt sporozoites. |
Liver stage | The sporozoites were found to invade and develop within HepG2 cells comparable to wt sporozoites. |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0403200 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0304600 | ||||||||||||||||||||||||||
Gene product | circumsporozoite (CS) protein | ||||||||||||||||||||||||||
Gene product: Alternative name | CSP | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | The repeat region of the endogenous gene is exchanged with the P. falciparum repeat region | ||||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | Not available | ||||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||||
Type of plasmid/construct | Plasmid single 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 | ||||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||||
Selection (positive) procedure | WR99210 | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | The mutant expresses a hybrid P. berghei/P. falciparum CS in which the P. berghei CS repeat region was exchanged with the P. falciparum CS repeat region. The P. falciparum CS repeat region was amplified from genomic DNA from the 7G8 isolate using primers that generated SpeI and XhoI sites: PfSpeI, 5'-GAGATGGAACTAGTAATAACGAAGACAACGAG-3'; PfXhoI, 5'-CTTGGTTCTTCCTCGAGGTTATTATTATTTTTTAC-3'. The CS repeat region within the CS gene of P. berghei in the construct used for transfection was exchanged with the P. falciparum CS repeat region using the SpeI and XhoI restriction sites | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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