RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5402

Malaria parasite	P. berghei
Genotype
Genetic modification not successful
Disrupted	Gene model (rodent): PBANKA_0922200; Gene model (P.falciparum): PF3D7_1126100; Gene product: autophagy-related protein 7, putative (Atg7)
Phenotype	No phenotype has been described

Last modified: 24 December 2024, 13:55

*RMgm-5402

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) : 39714137
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	Not applicable
Other information parent line	Plasmodium berghei ANKA (MRA 311) was obtained from BEI resources, USA
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Attempts to generate the mutant parasite were performed by
Name PI/Researcher	Mishra A, Mishra S
Name Group/Department	Division of Molecular Microbiology and Immunology
Name Institute	CSIR-Central Drug Research Institute
City	Lucknow
Country	India

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0922200

Gene Model P. falciparum ortholog

PF3D7_1126100

Gene product

autophagy-related protein 7, putative

Gene product: Alternative name

Atg7

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Details of the genetic modification

Inducable system used

Additional remarks inducable system

Type of plasmid/construct used

(Linear) plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used

Modified PlasmoGEM construct/vector used

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/yfcu

Promoter of the selectable marker

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth.

In an attempt to generate direct knockout of PbAtg7, two fragments, F3 (0.54 kb) and F4 (0.61 kb), were amplified using primers 1321/1322 and 1323/1324 and sequentially cloned and inserted into pBC-GFP-yFCU-hDHFR at blunt-ended SalI and NotI/AscI sites, respectively. The plasmid was linearized with XhoI/AscI restriction digestion and transfected into P. berghei ANKA schizonts

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

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