Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene tagging
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Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 25185663 |
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 2.34
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Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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The mutant parasite was generated by |
Name PI/Researcher | Tremp AZ; Dessens JT |
Name Group/Department | Pathogen Molecular Biology Department, Faculty of Infectious and Tropical Diseases |
Name Institute | London School of Hygiene and Tropical Medicine |
City | London |
Country | UK |
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Name of the mutant parasite |
RMgm number | RMgm-1121 |
Principal name | IMC1e/GFP |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype |
Asexual blood stage | Weak expression of IMC1e::GFP in blood stages |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Strong expression of IMC1e::GFP in blood stages that was distributed predominantly in the cell cortex, consistent with a pellicular localization of the protein. PbIMC1e was also present in an unknown structure situated at one extremity (posterior) of the ookinete. |
Oocyst | Sporulated oocysts and sporozoites also displayed GFP florescence, which localized to the periphery of the sporozoites. Sporozoites possessed a small fluorescent spot at one extremity (posterior). |
Sporozoite | Sporulated oocysts and sporozoites also displayed GFP florescence, which localized to the periphery of the sporozoites. Sporozoites possessed a small fluorescent spot at one extremity (posterior). |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of IMC1e (inner membrane complex protein 1e, putative).
Protein (function)
The three motile and invasive stages (zoites) of Plasmodium species (i.e. ookinetes, sporozoites and merozoites), as well as zoites of other apicomplexan parasites, possess a similar cortical structure termed the pellicle. The pellicle is essentially made up of the plasma membrane and an underlying double membrane structure termed the inner membrane complex (IMC). Closely associated with the IMC on its cytoplasmic side is a network of intermediate filaments termed the subpellicular network (SPN), which supports the pellicular membranes and provides mechanical strength to the cell. Several members of an Apicomplexa-specific family of proteins termed IMC1 proteins have been identified as components of the SPN. Structurally related proteins from ciliates and dinoflagellate algae have since been added to this protein family renamed ‘alveolins’, which now defines the Alveolata infrakingdom. In the genus Plasmodium, the number of members of the alveolin family has risen to 12, which are encoded by conserved and syntenic genes. The alveolin family members display differential expression between the three zoite stages of the parasite, with the largest repertoires present in the ookinete and sporozoite according to proteomic studies. It has been shown in the rodent malaria species Plasmodium berghei that the disruption of individual alveolin family members expressed in sporozoites (PbIMC1a), in ookinetes (PbIMC1b) or in both these zoites (PbIMC1h) results in morphological abnormalities that are accompanied by reduced tensile strength of the zoite stages in which they are expressed. Besides roles in morphogenesis and mechanical strength, the Plasmodium alveolins are also involved in gliding motility in both ookinetes and sporozoites, most likely through interactions with components of the glideosome that are situated within the pellicular cytoplasm.
PbIMC1e (PBANKA_040270) is composed of 512 amino acids encoded by a single exon. Sequence conservation is limited to an IMCp domain in their central portions. the Plasmodium imc1e locus is located directly downstream of its family member imc1a in the opposite orientation.
Phenotype
Unsuccessful attempts to to knock-out imc1e (RMgm-1119) indicates an essential role of IMC1e in blood stage development/multiplication.
GFP-tagging had no effect on the viability of all blood stages
Analysis of the mutant expressing GFP-tagged IMC1e showed:
Weak expression in blood stages whereas mature ookinetes displayed much stronger GFP fluorescence that was distributed predominantly in the cell cortex, consistent with a pellicular localization of the protein. PbIMC1e was also present in an unknown structure situated at one extremity (posterior) of the ookinete. Sporulated oocysts and sporozoites also displayed GFP florescence, which localized to the periphery of the sporozoites. Sporozoites possessed a small fluorescent spot at one extremity (posterior). Evidence is presented for temporal recruitment to the SPN of ookinetes.
Additional information
Other mutants
Unsuccessful attempts to to knock-out imc1e (RMgm-1119)
Click on IMC1 for other gene-deletion/tagging mutants targeting IMC1 proteins |