Back to search resultsSummaryRMgm-4979
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Successful modification | The 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) : 33665945 |
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 |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Zhenkui C; Yuan J |
Name Group/Department | State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, Schoo |
Name Institute | Xiamen University |
City | Xiamen, Fujian |
Country | China |
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Name of the mutant parasite | |
RMgm number | RMgm-4979 |
Principal name | Δap2-o3 |
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 | Normal numbers of gametocytes are produced. Normal exflagellation. Normal egress of gametes. Males are fertile (shown in crossing-experiments). Fertilization ability of females is impaired. Fertilized zygotes of the Δap2-o3 mutants were developmentally arrested at the early stages (stage II-III), displaying incomplete elongation. Only ookinetes with aberrant morphology are produced. |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation See mutant RMgm-4978 for expression analyses: Tagged AP2-O3 expressed in the nucleus of female gametocytes (not in males) and in mature oocysts (not in ookinetes, early stage oocysts and sporozoites). Evidence is presented that: |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_1017000 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1429200 | ||||||||||||||||||||||||
Gene product | AP2 domain transcription factor AP2-O3, putative | ||||||||||||||||||||||||
Gene product: Alternative name | ApiAP2; AP2-O3; PbAP2-FG2 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | 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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | CRISPR/Cas9 plasmid pYCm was used for genomic modification (Zhang et al, 2014; Zhang et al, 2017a). To construct the plasmids for gene deleting, the 50 - and 30-flanking genomic sequence (400–700 bp) of target genes was PCR-amplified as left and right homologous arms and inserted into the restriction sites of pYCm. Oligonucleotides for small guide RNAs (sgRNAs) were annealed and ligated into pYCm. Two sgRNAs were designed to target the coding region of each gene. | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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