RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5292
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1360100; Gene model (P.falciparum): PF3D7_1347200; Gene product: nucleoside transporter 1 (NT1)
Transgene
 Transgene not Plasmodium: mCherry
Promoter: Gene model: Not available; Gene model (P.falciparum): Not available; Gene product: Not available
3'UTR: Gene model: Not available; Gene product: Not available
Replacement locus: Gene model: PBANKA_1360100; Gene product: nucleoside transporter 1 (NT1)
Phenotype Asexual bloodstage;
Last modified: 6 March 2023, 10:06
  *RMgm-5292
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 36653222
MR4 number
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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 Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherDeveci G, Aly ASI
Name Group/DepartmentAly Lab, Beykoz Institute of Life Sciences and Biotechnology
Name InstituteBezmialem Vakif University
CityIstanbul
CountryTurkey
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Name of the mutant parasite
RMgm numberRMgm-5292
Principal namePbnt1(-)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stage(some) evidence presented for reduced blood stage growth/multiplication and reduced/absence of virulence
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of NT1 and expresses mCherrry

Protein (function)
Plasmodium lacks the enzymatic machinery necessary for the synthesis of the purine ring de novo and rely solely on the uptake of host purines for DNA synthesis. For P. falciparum it has been demonstrated that the translocation of host purines into the parasite is mainly mediated by the plasma membrane permease PfNT1 (nucleotide transporter 1). Genetic studies demonstrated that PfNT1 plays an essential role in parasite development and replication within human erythrocytes and parasites lacking PfNT1 are conditionally lethal, growing only when purines are provided at supra-physiological concentrations (El Bissati et al., 2006, PNAS 103, 9286-91; El Bissati, 2008, Mol. Biochem. Parasitol. 161, 130-9).

P. yoelii mutants (RMgm-387) and P. berghei mutants (RMgm-831; RMgm-836) lacking NT1 showed reduced blood stage growth/multiplication 

Phenotype
(some) evidence presented for reduced blood stage growth/multiplication and reduced/absence of virulence

See also other (published) P. yoelii mutants (RMgm-387) and P. berghei mutants (RMgm-831; RMgm-836) lacking NT1 that showed reduced blood stage growth/multiplication 

Additional information

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1360100
Gene Model P. falciparum ortholog PF3D7_1347200
Gene productnucleoside transporter 1
Gene product: Alternative nameNT1
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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 parasitehdhfr
Promoter of the selectable markerunknown
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
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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  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene namemCherry
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct(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
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markerunknown
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite Not available
Gene Model P. falciparum ortholog Not available
Gene productNot available
Gene product: Alternative name
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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3'-UTR
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative name
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PBANKA_1360100
Gene productnucleoside transporter 1
Gene product: Alternative nameNT1
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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
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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