RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5554
Malaria parasiteP. yoelii
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PYYM_1014900; Gene model (P.falciparum): PF3D7_1431400; Gene product: Surface-related antigen SRA (SRA)
PhenotypeNo phenotype has been described
Last modified: 7 October 2024, 12:34
  *RMgm-5554
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 38624215
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
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherFeng X, Cheng Y
Name Group/DepartmentDepartment of Public Health and Preventive Medicine, Laboratory of Pathogen Infection and Immunity,
Name InstituteWuxi School of Medicine, Jiangnan University
CityWuxi
CountryChina

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PYYM_1014900
Gene Model P. falciparum ortholog PF3D7_1431400
Gene productSurface-related antigen SRA
Gene product: Alternative nameSRA
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedCRISPR/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
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption Complete and partial disruption attempted
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth/multiplication

P. falciparum surface-related antigen (SRA) is located on the surface of merozoites and gametocytes, exported to the surface of infected erythrocyte membranes, and reported to bind to human erythrocytes.

The PySRA protein contains a signal peptide (aa 1–24) and a transmembrane domain (aa 853–871)

Different protein segements: PySRA-F1 (aa 24–298), PySRA-F2 (aa 299–548), and PySRA-F3 (aa 705–852)

Complete PySRA and PySRA-f2 could not be knocked out/disrupted

From the Abstract:
This study found that the F2 segment of P. yoelii SRA activated downstream MAPK and NF-κB signaling pathways by binding to CD68 on the surface of the macrophage membrane and regulating the inflammatory response. The anti-PySRA-F2 antibody can protect mice against P. yoelii, and the pro-inflammatory responses such as IL-1β, TNF-α, and IL-6 after infection with P. yoelii are attenuated.
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