RMgmDB - Rodent Malaria genetically modified Parasites
Back to search resultsSummaryRMgm-1475
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Last modified: 5 July 2016, 17:10
*RMgm-1475| 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 | 3 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 27353755 |
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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 | Not applicable |
| Other information parent line | |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Voss C; Sinnis P; Coppens I |
| Name Group/Department | Department of Molecular Microbiology and Immunology, Malaria Research Institute |
| Name Institute | Johns Hopkins University Bloomberg School of Public Health |
| City | Baltimore, Maryland |
| Country | USA |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0504100 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1019900 | ||||||||||||||||||||||||
| Gene product | autophagy-related protein 8 | ||||||||||||||||||||||||
| Gene product: Alternative name | ATG8 | ||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | unknown | ||||||||||||||||||||||||
| 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 | Unknown | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | unknown | ||||||||||||||||||||||||
| Promoter of the selectable marker | unknown | ||||||||||||||||||||||||
| Selection (positive) procedure | unknown | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | The negative attempts to disrupt the atg8 gene indicates an essential role of ATG8 for blood stage development/multiplication. no details are provided in the paper for the construct used in attempts to disrupt the gene. See also RMgm-1476 for a P. berghei mutant over-expressing ATG8. Autophagy is the archetypal disposal pathway for keeping the cell interior clean of disused and superfluous organelles In contrast to mammalian cells and yeast that contain ~40 autophagy-related genes (ATG), the malaria parasite contains ~15 orthologs of ATG identified by in silico comparative studies: those whose products are required for vesicle expansion and completion are present, while genes involved in induction of autophagy and cargo packaging are mostly absent. All the components of the ubiquitin-like ATG8 system, which are involved in autophagosome formation, are expressed by Plasmodium. Coexpression of PbATG8, PbATG3, and PbATG7 is upregulated during early sporozoite differentiation in liver cells. In liver forms of P. berghei, PbATG8 localizes to a network of vesicles and tubules that align along the apicoplast, a relict plastid organelle, as well as to numerous vesicles in the cytoplasm. PbATG8 is preferentially distributed on the outermost membranes of the apicoplast. ATG8 expressed in the related apicomplexan parasite Toxoplasma gondii is also localized to the apicoplast, and it seems to be involved in the maintenance of this organelle and proper segregation between daughter cells during replication. A mutant over-expressing PbATG8 (RMgm-1476) showed poor development into late liver stages and production of small merosomes that contain immature merozoites unable to initiate a blood infection. Based on these observations it has been proposed that that Plasmodium ATG8 is a key effector in the development of merozoites by controlling microneme clearance and apicoplast proliferation and that dysregulation in ATG8 levels is detrimental for malaria infectivity. | ||||||||||||||||||||||||
| 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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