RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1453
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene Plasmodium: Gene model: PBANKA_0208000; Gene model (P.falciparum): PF3D7_0105300; Gene product: cyclase-associated protein (CAP; C-CAP)
Promoter: Gene model: PBANKA_0403200; Gene model (P.falciparum): PF3D7_0304600; Gene product: circumsporozoite (CS) protein (CSP)
3'UTR: Gene model: PBANKA_0208000; Gene product: cyclase-associated protein (CAP; C-CAP)
Insertion locus: Gene model: PBANKA_0208000; Gene product: cyclase-associated protein (CAP; C-CAP)
Phenotype Sporozoite; Liver stage;
Last modified: 17 June 2016, 18:57
  *RMgm-1453
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 27226484
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/ResearcherSato Y; Matuschewski K
Name Group/DepartmentParasitology Unit
Name InstituteMax Planck Institute for Infection Biology
CityBerlin
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-1453
Principal nameC-CAP(CSP)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteStrong reduction in salivary gland sporozoites. Sporozoite motility affected (see below).
Liver stageReduced in vitro invasion of hepatocytes by C-CAP(CSP) sporozoites (but normal prepatent period after mosquito infection of mice)
Additional remarks phenotype

Mutant/mutation
The mutant contains an additional copy of the c-cap gene under the control of the csp promoter. The c-cap gene is N-terminal tagged with a FLAG tag

Protein (function)
Cyclase associated proteins (CAP) are involved in the turnover of actin (F-actin) cytoskeletal structures in part through binding to monomeric actin (G-actin). The Plasmodium protein consists of the conserved actin binding domain and lacks the adenylate cyclase binding domain and the WH2 domain.

Phenotype
In this study three mutants are generated and analysed that contain an additional copy of the c-cap gene under the control of three different promoters (ama-1, ctrp and csp). See also mutants RMgm-1451 and RMgm-1454

No phenotype was observed in blood stages

The C-CAP(CTRP) produced normal numbers of ookinetes with normal motility and normal oocyst production

In the C-CAP(AMA1) mutant a strong reduction in salivary gland sporozoites was observed (2,000-5,000 salivary gland sporozoites, a two- to five-fold reduction as compared to WT sporozoites).
In the  C-CAP(CSP) very few salivary gland sporozoites were observed.
Sporozoite motility: C-CAP(CSP) sporozoites displayed either incomplete or full trails, but of only few (<10) circles. C-CAP(AMA1) sporozoites showed a reduced number of trails compared to WT, but the majority was able to perform productive motility.
Reduced in vitro invasion of hepatocytes by both C-CAP(CSP) and C-CAP(AMA1) sporozoites (but normal prepatent period after mosquito infection of mice).

Additional information

Other mutants
See PF3D7_0105300 for other 'cyclase-associated' mutants

  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: Plasmodium
Gene Model of Parasite PBANKA_0208000
Gene Model P. falciparum ortholog PF3D7_0105300
Gene productcyclase-associated protein
Gene product: Alternative nameCAP; C-CAP
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct(Linear) plasmid single 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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe csp/c-cap construct was integrated into the c-cap gene locus by single cross-over integration resulting in allele duplication with one WT c-cap copy and one copy under the control of the stage-specific promoter.
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_0403200
Gene Model P. falciparum ortholog PF3D7_0304600
Gene productcircumsporozoite (CS) protein
Gene product: Alternative nameCSP
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 PBANKA_0208000
Gene productcyclase-associated protein
Gene product: Alternative nameCAP; C-CAP
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 / InsertionInsertion locus
Gene Model of Parasite PBANKA_0208000
Gene productcyclase-associated protein
Gene product: Alternative nameCAP; C-CAP
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