SummaryRMgm-5168
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Successful modification | The 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) : 35775739 |
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 | Quian P, Yuan J |
Name Group/Department | State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signal Network, School o |
Name Institute | Xiamen University |
City | Xiamen |
Country | China |
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Name of the mutant parasite | |
RMgm number | RMgm-5168 |
Principal name | see below |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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Phenotype | |
Asexual blood stage | IMC location in blood stages |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Among these proteins, the orthologues of 8 proteins have been experimentally validated to be IMC-residing or association in the schizonts of P. berghei or P. falciparum,
PY17X_ 0314700 (CDPK1), PY17X_0617900 (CDPK4), PY17X_1440500 (PKAr), PY17X_0839000 (PKAc),
PY17X_1420600 (Rab11A), PY17X_1462100 (MTIP), PY17X_0206000 (PhIL1), PY17X_0525300 (GAPM2) IMC localization or association of the 14 other candidates have not been well characterized in the Plasmodium PY17X_0207400 (no IMC location) PY17X_0312400 (IMC) PY17X_0417300 (no IMC location) PY17X_0418000 (IMC) PY17X_0812700 (IMC)
PY17X_0917100 (no IMC location) PY17X_1131200 (IMC) PY17X_1139700 (IMC) PY17X_1220300 (IMC) PY17X_1348200 (IMC) PY17X_1359500 (IMC) PY17X_1411000 (IMC) PY17X_1441500 (IMC) PY17X_1453100 (IMC). Immunoblot assays were used to detect each protein, all displaying a band fitting their expected molecular weight. As expected, immunofluorescence assays (IFA) showed clear co-localization of the 8 known proteins (CDPK1, CDPK4, PKAr, PKAc, Rab11A, MTIP, PhIL1, 247 and GAPM2) with the IMC marker GAP45. Among the 14 newly characterized candidates, 11 of them (PY17X_0312400, PY17X_0418000, PY17X_0812700, PY17X_1131200, PY17X_1139700, PY17X_1220300, PY17X_1348200, PY17X_1359500, PY17X_1411000, PY17X_1441500, and PY17X_1453100) displayed the IMC or IMC-like pellicle localization, while 3 other candidates (PY17X_0207400, PY17X_0417300 and PY17X_0917100) did not. Collectively, we confirmed the IMC or IMC-like localization of 19 proteins from the 22 candidates in the P. yoelii schizonts. Additional information Other mutants |
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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: Plasmodium | ||||||||||||||||||
Gene Model of Parasite | PY17X_0812700 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0908000 | ||||||||||||||||||
Gene product | plasma membrane protein 1, putative | ||||||||||||||||||
Gene product: Alternative name | PMP1 | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||
Type of plasmid/construct | Circular plasmid | ||||||||||||||||||
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 | No | ||||||||||||||||||
Additional remarks genetic modification | CRISPR/Cas9 plasmid pYCm was used for parasite genomic modification. To construct the plasmids for gene tagging, the DNA fragment encoding 6HA was inserted between the left and right arms in frame with the gene of interest. For each gene tagging, two sgRNAs were designed to target sites close to the N- or C-terminal part of the coding region. Each candidate gene was tagged with a 6HA at the N- or C-terminus and driven by the promoter of gene isp3 (PBANKA_1324300) for episomal expression in the asexual blood stages. | ||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PBANKA_1324300 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1460600 | ||||||||||||||||||
Gene product | inner membrane complex sub-compartment protein 3 | ||||||||||||||||||
Gene product: Alternative name | ISP3 | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | Not available | ||||||||||||||||||
Gene product | Not available | ||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Not available | ||||||||||||||||||
Gene Model of Parasite | Not available | ||||||||||||||||||
Gene product | Not available | ||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||
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