Back to search resultsSummaryRMgm-4592
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Successful modification | The 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 | 3 |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 30703164 |
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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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Attempts to generate the mutant parasite were performed by | |
Name PI/Researcher | Hart KJ, Lindner SE |
Name Group/Department | Department of Biochemistry and Molecular Biology, Center for Malaria Research |
Name Institute | Pennsylvania State University, University Park, State College |
City | Pennsylvania |
Country | USA |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_1428300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0811300 | ||||||||||||||||||||||||
Gene product | CCR4-associated factor 1 | ||||||||||||||||||||||||
Gene product: Alternative name | CAF1 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) plasmid double cross-over | ||||||||||||||||||||||||
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 | unknown | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | Multiple attempts to delete the caf1 gene indicate an essential function for growth/multiplication of asexual blood stages. In addition to transcript stabilization, translational control can also be accomplished by the degradation of transcripts. Degradation of mRNAs is typically initiated by deadenylases, which remove the protective poly(A) tail. In many eukaryotes, the main complex responsible for deadenylation is the CAF1/CCR4/NOT complex, which also participates in transcriptional elongation, translational repression, and histone modification functions, and thus acts broadly upon gene expression. To first assess the importance of the CAF1/CCR4/NOT complex in Plasmodium, we bioinformatically identified the genes for all members of the canonical CAF1/CCR4/NOT complex in Plasmodium, except for not3 and caf130. The absence of these two particular genes is not surprising, as these genes are also absent in some eukaryotes. In addition, we identified four CCR4 domain-containing proteins (PyCCR4-1, PyCCR4-2, PyCCR4-3, PyCCR4-4) that have homology to CCR4 deadenylases in other eukaryotes (e.g. yeast, human, mouse). | ||||||||||||||||||||||||
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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