SummaryRMgm-5284
|
Successful modification | The 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) : 36463257 |
MR4 number | |
top of page | |
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 |
Other information parent line | |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | Quian P, 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 |
Country | China |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-5284 |
Principal name | apr2::gfp;ara1::6HA |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page | |
Phenotype | |
Asexual blood stage | Not tested |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Close association between APR2 and ARA1 |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation The mutant expresses ARA1 C-terminally tagged with 6HA. In addition, it expresses APR2 tagged with GFP at the C-terminus. The endogenous ARA1, APRp2 and APRp4 proteins were tagged with a 6HA using the CRISPR-Cas9 in apr2::gfp background (RMgm-5282) generating 3 single-tagged strains 3 double-tagged strains (apr2::gfp;ara1::6HA, apr2::gfp;aprp2::6HA, and apr2::gfp;aprp4::6HA) (see RMgm-5284, RMgm-5285, RMgm-5286). Protein (function) Phenotype Additional information Other mutants |
top of page | |||||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_1339500 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1471600 | ||||||||||||||||||||||||||
Gene product | apical polar ring protein APR2, putative | ||||||||||||||||||||||||||
Gene product: Alternative name | APR2 | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | GFP | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | We tagged the APR2 with GFP at either the N- or C-terminus in the 17XNL parasite using the CRISPR-Cas9 method, generating GFPA::apr2 and apr2::GFP clones | ||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | CRISPR/Cas9 construct: integration through double strand break repair | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | We tagged the APR2 with GFP at the C-terminus in the 17XNL parasite using the CRISPR-Cas9 method, generating GFP::apr2 and apr2::GFP clones | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |
top of page | |||||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_1412750 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1312450 | ||||||||||||||||||||||||||
Gene product | apical ring associated protein 1, putative | ||||||||||||||||||||||||||
Gene product: Alternative name | ARA1 | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | 6xHA | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | CRISPR/Cas9 construct: integration through double strand break repair | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | We tagged the ARA1 with 6HA at the C-terminus. The endogenous ARA1, APRp2 and APRp4 proteins were tagged with a 6HA using the CRISPR-Cas9 in both WT and apr2::gfp background generating 3 single-tagged strains (ara1::6HA, aprp2::6HA, and aprp4::6HA) and 3 double-tagged strains (apr2::gfp;ara1::6HA, apr2::gfp;aprp2::6HA, and apr2::gfp;aprp4::6HA) (see RMgm-5284, RMgm-5285, RMgm-5286). | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |