RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4477
Malaria parasiteP. yoelii
Genotype
TaggedGene model (rodent): PY17X_1323300; Gene model (P.falciparum): PF3D7_1455800; Gene product: LCCL domain-containing protein (CCp2, LAP4)
Name tag: mCherry
TaggedGene model (rodent): PY17X_0418900; Gene model (P.falciparum): PF3D7_0905300; Gene product: dynein heavy chain, putative (Dhc1)
Name tag: GFP
Phenotype Gametocyte/Gamete;
Last modified: 15 August 2018, 16:12
  *RMgm-4477
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging, Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 28034675
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherLiu, C; Yuan, J
Name Group/DepartmentState Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Biology, School of Life
Name InstituteXiamen University
CityXiamen, Fujian
CountryChina
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Name of the mutant parasite
RMgm numberRMgm-4477
Principal nameCCp2::mCherry/Dhc1::GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteCCp2::mCherry expression in female gametocytes
Dhc1::GFP expression in male gametocytes/gametes
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal mCherry-tagged version of CCp2 and a C-terminal GFP-tagged version of Dhc1. The mutant does not contain a drug-selectable marker.

This mutant was generated by crossing of a mutant expressing mCherry-tagged CCp2 in female gametocytes (RMgm-4475) with a mutant expressing GFP-tagged Dhc1 in male gametocytes (RMgm-4476),

For generation of the mutants expressing mCherry-tagged CCp2 and GFP-tagged Dhc1 the CRISPR/Cas9 method was used as described for mutants RMgm-1095 and RMgm-4071.

Protein (function)
The lap4 (ccp2) gene is a member of a small conserved gene family (6 genes), encoding proteins with multiple adhesive domains, for example a Lgl1 (LCCL)-lectin adhesive domain. In P. falciparum the LCCL domain-containing proteins are termed PfCCp's and in P. berghei PbLAP's. CCp2 is a female specific protein, transcribed/expressed in female gametocytes, ookinetes and young gametocytes

Dhc1 is a a male gametocyte/gamete-specific dynein heavy chain protein

Phenotype
CCp2::mCherry expression in female gametocytes
Dhc1::GFP expression in male gametocytes/gametes

Additional information
In P. berghei transcripts of ccp2 are under translational control in female gametocytes. The protein is only expressed after activation of gametocytes (in gametes/zygotes/ookinetes). In the mutants described here the mCherry-tagged version of P. yoelii ccp2 is expressed as a protein in female gametocytes.

Other mutants

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_1323300
Gene Model P. falciparum ortholog PF3D7_1455800
Gene productLCCL domain-containing protein
Gene product: Alternative nameCCp2, LAP4
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Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/constructCRISPR/Cas9 construct: integration through double strand break repair
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/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modificationFor tagging CCp2 the CRISPR/Cas9 method was used as described for mutants RMgm-1095 and RMgm-4071.
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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  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_0418900
Gene Model P. falciparum ortholog PF3D7_0905300
Gene productdynein heavy chain, putative
Gene product: Alternative nameDhc1
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Details of the genetic modification
Name of the tagGFP
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/constructCRISPR/Cas9 construct: integration through double strand break repair
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/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modificationFor tagging Dhc1 the CRISPR/Cas9 method was used as described for mutants RMgm-1095 and RMgm-4071.
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: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren