RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-4994
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Last modified: 18 May 2021, 18:00
*RMgm-4994| Successful modification | The gene/parasite could not be changed/generated by the genetic modification. |
| The following genetic modifications were attempted | Gene disruption |
| Number of attempts to introduce the genetic modification | 4 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 33762339 |
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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 |
| 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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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Balestra AC, Brochet M |
| Name Group/Department | Department of Microbiology and Molecular Medicine, Faculty of Medicine |
| Name Institute | University of Geneva |
| City | Geneva |
| Country | Switzerland |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1446100 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | Pf3D7_ 1231400 | ||||||||||||||||||||||||
| Gene product | membrane protein ICM1 | ||||||||||||||||||||||||
| Gene product: Alternative name | ICM1 | ||||||||||||||||||||||||
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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 | Yes | ||||||||||||||||||||||||
| Name of PlasmoGEM construct/vector | PbGEM-265644 | ||||||||||||||||||||||||
| 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 | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood-stage growth/multiplication. From the paper: 'We identify a multipass membrane protein, termed important for calcium mobilization-1 (ICM1; Pf3D7_1231400) that interacts with and is phosphorylated by PKG in two different Plasmodium developmental stages and species. Through stage-specific knockdown and reverse genetic approaches, we show that ICM1 is essential for calcium mobilization in both the clinically relevant asexual blood stages and in gametocytes that mediate transmission to mosquitoes. Our findings highlight this putative transporter or channel as a crucial link between PKG function and calcium signaling'. PKG is involved in release of merozoites from red blood cells and hepatocytes, induction of gametogenesis and parasite transmission upon ingestion of gametocytes by a blood-feeding mosquito, and sustaining cellular motility necessary for parasite dissemination. A major function of PKG in all these processes is the tightly regulated mobilization of calcium from intracellular stores within seconds of activation In the paper evidence is presented that: - ICM1 may represent a structurally divergent polytopic membrane protein with architectural similarities to mammalian channels, transporters, and IP3 receptors. - ICM1 is phosphorylated in a PKG-dependent manner - Stage-specific knockdown of PbICM1 reveals a crucial role in early calcium mobilization required to initiate gametogenesis | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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