RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-4093
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Last modified: 15 January 2017, 12:57
*RMgm-4093| 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) : 28081440 |
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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 676m1cl1 (RMgm-29) |
| Other information parent line | 676m1cl1 (RMgm-29) is a reference ANKA mutant line which expresses GFP-luciferase under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190). |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Modrzynska K, Billker O |
| Name Group/Department | Wellcome Trust Sanger Institute |
| Name Institute | Wellcome Trust Sanger Institute |
| City | Hinxton, Cambridge |
| Country | UK |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1415700 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1317200 | ||||||||||||||||||||||||
| Gene product | AP2 domain transcription factor, putative | ||||||||||||||||||||||||
| Gene product: Alternative name | ApiAP2 | ||||||||||||||||||||||||
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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 | - | ||||||||||||||||||||||||
| 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 delete this gene indicate an essential function during asexual blood stage development From the paper: To target P. berghei ApiAP2 genes systematically, we succeeded in producing deletion vectors for all but one member of the family (PBANKA_1313200) and transfected each of them into a reporter line constitutively expressing GFP to facilitate phenotyping. Fourteen ApiAP2 genes resisted at least four disruption attempts with up to two different vector designs, providing tentative evidence that more than half of the genes in this family are potentially essential for asexual blood stage growth in vivo. For the remaining eleven, a KO line could be generated. These included six genes that had not previously been studied in Plasmodium. Most sequence-specific transcription factor families found in other eukaryotes seem to be absent from Plasmodium. Instead, an expansion of a protein family containing one or more apetala2 (AP2) DNA-binding domains was observed across the phylum apicomplexa. In total, 27 members of this family have been found in the human malaria parasite Plasmodium falciparum (although a possible 28th member of the family may be present. In total, 26 of these have syntenic orthologs in rodent malaria species, each with its unique stage-specific expression profile. The paper presents a systematic knockout (KO) screen targeting the ApiAP2 family in the rodent malaria parasite P. berghei. Phenotyping of eleven viable ApiAP2 KO mutants reveals ten critical gene functions at different points in the life cycle. | ||||||||||||||||||||||||
| 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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