Summary

RMgm-4405
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1434600; Gene model (P.falciparum): PF3D7_1216600; Gene product: cell traversal protein for ookinetes and sporozoites (CelTOS, S4)
Phenotype Fertilization and ookinete; Oocyst; Sporozoite;
Last modified: 1 January 2018, 19:31
  *RMgm-4405
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 29253313
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherSteel RWJ, Kappe SHI
Name Group/DepartmentCenter for Infectious Disease Research
Name Instituteformerly Seattle Biomedical Research Institute
CitySeattle
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-4405
Principal namePy s4/celtos(-)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNormal gametocyte and ookinete production. Surprisingly PPy s4/celtos¯ parasites also established a low-level mosquito midgut infection.
OocystNormal gametocyte and ookinete production. Surprisingly Py s4/celtos¯ parasites also established a low-level mosquito midgut infection. Whereas the number of oocysts per midgut stayed stable between days 7 – 11 for Py WT, oocyst numbers for Py s4/celtos¯ parasites declined throughout this period. The fraction of midguts infected with Py s4/celtos¯ declined markedly from day 7 - 11, by which time oocysts were only occasionally observed and these exhibited defects in sporulation compared to Py WT oocysts. Combined, the data suggest that Py S4/CelTOS has an important role during oocyst survival and/or sporogenesis in addition to the previously reported role in ookinete traversal of the midgut epithelium.
SporozoiteNo salivary gland sporozoites.
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of CelTOS (S4)

Protein (function)
CelTOS is localized to the micronemes of ookinetes and (salivary gland) sporozoites and is secreted. It plays a role in migration of ookinetes through mosquito midgut cells and traversal of sporozoites through heptocytes

Phenotype
Normal gametocyte and ookinete production. Surprisingly Py s4/celtos¯ parasites also established a low-level mosquito midgut infection. Whereas the number of oocysts per midgut stayed stable between days 7 – 11 for  Py WT, oocyst numbers for Py s4/celtos¯ parasites declined throughout this period. The fraction of midguts infected with Py s4/celtos¯ declined markedly from day 7 - 11, by which time oocysts were only occasionally observed and these exhibited defects in sporulation compared to Py WT oocysts. Combined, the data suggest that Py S4/CelTOS has an important role during oocyst survival and/or sporogenesis in addition to the previously reported role in ookinete traversal of the midgut epithelium. No salivary gland sporozoites.

Additional information

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_1434600
Gene Model P. falciparum ortholog PF3D7_1216600
Gene productcell traversal protein for ookinetes and sporozoites
Gene product: Alternative nameCelTOS, S4
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
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
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationTo generate Py s4/celtos¯, two DNA fragments containing approximately 800 bp of the 5′ UTR and 3′ UTR of Py S4/CelTOS were amplified from WT Py 17XNL genomic DNA and cloned into the pDEF vector (BEI Resources).
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