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Details of the target gene |
Gene Model of Rodent Parasite |
PY17X_1241800
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Gene Model P. falciparum ortholog |
PF3D7_0523800
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Gene product | food vacuole resident transporter 1 |
Gene product: Alternative name | FVRT1, PyDMT1 (Divalent metal transporter 1) |
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Details of the genetic modification |
Inducable system used | No |
Additional remarks inducable system |
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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
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Plasmid/construct map |
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Plasmid/construct sequence |
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Restriction sites to linearize plasmid |
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Partial or complete disruption of the gene | Complete |
Additional remarks partial/complete disruption |
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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 | The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth/multiplication
To clarify the physiological function of PyDMT1 in Plasmodium, we attempted to knock out PyDMT1 in P. yoelii17X by CRISPR-Cas9. Nine guide RNAs were designed to cover the Pydmt1 gene, and each guide RNA and homologous recombination arms were independently electro-transfected five times. No Pydmt1 knockout parasites were obtained in all these trials. |
Additional remarks selection procedure | |
Primer information: Primers used for amplification of the target sequences 
Primer information: Primers used for amplification of the target sequences
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 | |
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