| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
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| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 34301597
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| MR4 number |
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| Parent parasite used to introduce the genetic modification |
| Rodent Malaria Parasite | P. yoelii |
| Parent strain/line | P. y. yoelii 17XNL |
| Name parent line/clone |
Not applicable
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| Other information parent line | |
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| The mutant parasite was generated by |
| Name PI/Researcher | Yang Z, Yuan J |
| Name Group/Department | State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signal Network, School o |
| Name Institute | Xiamen University |
| City | Xiamen, Fujian |
| Country | China |
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| Name of the mutant parasite |
| RMgm number | RMgm-5053 |
| Principal name | soap-2a-mCherry |
| Alternative name | OmCherry |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | No |
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| Phenotype |
| Asexual blood stage | Not tested |
| Gametocyte/Gamete | Not different from wild type |
| Fertilization and ookinete | Since the soap gene is highly expressed in mature ookinetes and early oocysts, we engineered two parasite strains Ogfp and OmCherry, wherein the endogenous soap sequence is fused with the coding sequence of green fluorescent protein (GFP) and mCherry, respectively, at the C terminus to track early oocysts. A “ribosome skip” T2A peptide was inserted between the SOAP and GFP or mCherry to ensure the separate expression of GFP or mCherry and the endogenous SOAP. As expected, GFP and mCherry fluorescence was detected specifically in the mature ookinetes in vitro and in the ookinetes and early oocysts in vivo |
| Oocyst | Since the soap gene is highly expressed in mature ookinetes and early oocysts, we engineered two parasite strains Ogfp and OmCherry, wherein the endogenous soap sequence is fused with the coding sequence of green fluorescent protein (GFP) and mCherry, respectively, at the C terminus to track early oocysts. A “ribosome skip” T2A peptide was inserted between the SOAP and GFP or mCherry to ensure the separate expression of GFP or mCherry and the endogenous SOAP. As expected, GFP and mCherry fluorescence was detected specifically in the mature ookinetes in vitro and in the ookinetes and early oocysts in vivo |
| Sporozoite | Not tested |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal 2A-GFP-tagged form of SOAP.
Since the soap gene is highly expressed in mature ookinetes and early oocysts, we engineered two parasite strains Ogfp and OmCherry, wherein the endogenous soap sequence is fused with the coding sequence of green fluorescent protein (GFP) and mCherry, respectively, at the C terminus to track early oocysts. A “ribosome skip” T2A peptide was inserted between the SOAP and GFP or mCherry to ensure the separate expression of GFP or mCherry and the endogenous SOAP. As expected, GFP and mCherry fluorescence was detected specifically in the mature ookinetes in vitro and in the ookinetes and early oocysts in vivo
The mutant has been generated using CRISPR/cas9 genome editing (using constructs as described for mutant RMgm-1095
Protein (function)
SOAP, a 21-kDa protein, has the hallmarks of an extracellular soluble (secreted) protein: it contains a predicted cleavable amino-terminal signal peptide, but lacks other typical organellar targeting or membrane anchoring signals. The conserved protein contains 12, closely spaced, cysteine residues and comparison of SOAP of different Plasmodium species indicates a modular structure of two cysteine-rich domains separated by a spacer sequence of variable amino acid length and composition.
SOAP is located in the micronemes of ookinetes.
Phenotype
Since the soap gene is highly expressed in mature ookinetes and early oocysts, we engineered two parasite strains Ogfp and OmCherry, wherein the endogenous soap sequence is fused with the coding sequence of green fluorescent protein (GFP) and mCherry, respectively, at the C terminus to track early oocysts. A “ribosome skip” T2A peptide was inserted between the SOAP and GFP or mCherry to ensure the separate expression of GFP or mCherry and the endogenous SOAP. As expected, GFP and mCherry fluorescence was detected specifically in the mature ookinetes in vitro and in the ookinetes and early oocysts in vivo
Additional information
Other mutants
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*RMgm-5053
