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) : 39037752 |
MR4 number |
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Parent parasite used to introduce the genetic modification |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
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 | Ghosh A, Mishra S |
Name Group/Department | Division of Molecular Microbiology and Immunology |
Name Institute | CSIR-Central Drug Research Institute |
City | Lucknow |
Country | India |
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Name of the mutant parasite |
RMgm number | RMgm-5589 |
Principal name | Scot1-3XHA-mCherry |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Expression of Scot1 in oocysts. Expression was not detected in the blood, gamete, ookinete. |
Sporozoite | Expression of Scot1 in oocysts and sporozoites. Expression was not detected in the blood, gamete, ookinete.
Sporozoites were stained with anti-mCherry and anti-TRAP antibodies. A granular localization pattern of Scot1-3XHA-mCherry was observed, which colocalized with the TRAP signal in sporozoites, indicating that Scot1 is a micronemal protein. |
Liver stage | Expression of Scot1 in liver stages (at 12, 24 and 62 hpi). Expression was not detected in liver stages at 36 or 48 hpi. |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal 3xHA-mCherry-tagged version of Scot1
Protein (function)
The Scot1 (micronemal) protein lacks a signal sequence and transmembrane domain
Phenotype
Expression of Scot1 in oocysts, sporozoites and liver stages (at 12, 24 and 62 hpi). Expression was not detected in the blood, gamete, ookinete, or liver stages (at 36 or 48 hpi). The localization of Scot1-3XHA-mCherry was analyzed in sporozoites and liver stages. Sporozoites were stained with anti-mCherry and anti-TRAP antibodies. A granular localization pattern of Scot1-3XHA-mCherry was observed, which colocalized with the TRAP signal in sporozoites, indicating that Scot1 is a micronemal protein.
Analysis of a mutant lacking expression of Scot1 (RMgm-5587) showed the following: Normal blood, oocyst and sporozoite production and normal invasion of hepatocytes by sporozoites. No blood stage infection in C57BL/6after after intravenous injection of salivary gland sporozoites or infected by mosquito bites. Normal invasion of hepatocytes. In livers of infected mice collected at 40 and 55 hpi, the parasite burden (quantified by amplifying 18S rRNA using real-time PCR), no difference in the 18S rRNA copy number was found at 40 hpi, but it was significantly lower at 55 hpi. Normal growth, numbers and sizes of liver stages. Reduced nuclear division at 62 hpi. No detached liver stages detected.
Additional information
Other mutants |