RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5251
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1365100; Gene model (P.falciparum): PF3D7_1352400; Gene product: conserved Plasmodium protein, unknown function (NUP176)
PhenotypeNo phenotype has been described
Last modified: 23 September 2022, 14:56
  *RMgm-5251
Successful modificationThe 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) : 36040030
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. 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/ResearcherAmbekar, SV, Mair GR
Name Group/DepartmentBiomedical Sciences,
Name InstituteIowa State University
CityAmes
CountryUSA
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1365100
Gene Model P. falciparum ortholog PF3D7_1352400
Gene productconserved Plasmodium protein, unknown function
Gene product: Alternative nameNUP176
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasiteunknown
Promoter of the selectable markerunknown
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe unsuccessful attempts to delete the gene indicate an essential role during asexual blood stage growth/multiplication

In the paper different genes are identified that encode putative nucleoporins (NUPs).
Two of the 5 novel, putative Nups are predicted to be essential for asexual life cycle progression from the large-scale PlasmoGEM forward mutagenesis screen in P. berghei while the remaining 3 (Nup335, Nup176, Nup269) were not targeted in this screen. In order to provide additional evidence for a critical role in parasite biology, we attempted to generate knockout lines for these 3 Nups using established methodologies Knockouts of Nup335, Nup176, and Nup269 failed repeatedly (n = 3 transfections for each gene), consistent with an important or essential role for these and all known Nups in the P. berghei blood stage in maintaining a functional NPC.
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren