RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1200

Malaria parasite	P. berghei
Genotype
Other modification	Gene model (rodent): PBANKA_0615200; Gene model (P.falciparum): PF3D7_0717500; Gene product: calcium-dependent protein kinase 4 (CDPK4) Details modification: Successful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants
Phenotype	No phenotype has been described

Last modified: 21 March 2015, 20:40

*RMgm-1200

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Other
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 25732065
MR4 number
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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
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The mutant parasite was generated by
Name PI/Researcher	Gomes AR; Billker O
Name Group/Department	Wellcome Trust Sanger Institute
Name Institute	Wellcome Trust Sanger Institute
City	Hinxton Cambridge
Country	UK
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Name of the mutant parasite
RMgm number	RMgm-1200
Principal name	-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Not different from wild type
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Successful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants. No evidence was found for reduced fitness (growth rate) of blood stages In this study P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest. In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants. In this study parasites were (mainly) transfected with a pool of vectors targeting protein kinase genes for deletion. Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis). To analyze growth curves derived from barcode counting two parameters were considered: (1) the relative abundance of each barcode within the pool, and (2) the relative fitness of each mutant, i.e., the rate at which its abundance changed each day (i.e. growth rate).

Other: Mutant parasite with another genetic modification

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Details of the target gene
Gene Model of Rodent Parasite	PBANKA_0615200
Gene Model P. falciparum ortholog	PF3D7_0717500
Gene product	calcium-dependent protein kinase 4
Gene product: Alternative name	CDPK4
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Description
Short description of the modification	Successful gene deletion as determined by barcode PCR in a large pool of gene-deletion mutants
Description	In this study P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest. In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants. In this study parasites were (mainly) transfected with a pool of vectors targeting protein kinase genes for deletion. Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis). The gene was targeted by PlasmoGEM vector PbGEM-087803
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