Back to search resultsSummaryRMgm-1346
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 26607162 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. yoelii |
Parent strain/line | P. y. yoelii 17XNL |
Name parent line/clone | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Risco-Castillo V; Silvie, O |
Name Group/Department | Centre d’Immunologie et des Maladies Infectieuses |
Name Institute | INSERM |
City | Paris |
Country | France |
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Name of the mutant parasite | |
RMgm number | RMgm-1346 |
Principal name | PyΔplp1 |
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 | Not different from wild type |
Sporozoite | Mutant sporozoites were motile and developed into EEFs in vitro (in HepG2/CD81 cells) as efficiently as control parasites, but were poorly infective to mice in vivo, especially when administered through mosquito bites, the natural transmission route. |
Liver stage | Mutant sporozoites were motile and developed into EEFs in vitro (in HepG2/CD81 cells) as efficiently as control parasites, but were poorly infective to mice in vivo, especially when administered through mosquito bites, the natural transmission route. |
Additional remarks phenotype | Mutant/mutation The mutant parasites are selected by a combination of positive selection (pyrimethamine), negative selection (5-FC) and FACS sorting (GOMO transfection; 'Gene Out Marker Out'; see mutant RMgm-1026 for more details for this transfection method and the construct used). |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_1007700 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0408700 | ||||||||||||||||||||||||
Gene product | perforin-like protein 1 | sporozoite micronemal protein essential for cell traversal | ||||||||||||||||||||||||
Gene product: Alternative name | PLP1; PPLP1, SPECT2 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) 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 FACS 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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