RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4828

Malaria parasite	P. berghei
Genotype
Mutated	Gene model (rodent): PBANKA_0208800; Gene model (P.falciparum): PF3D7_0104300; Gene product: ubiquitin carboxyl-terminal hydrolase 1, putative (UBP1) Details mutation: UBP1 mutated - V2721F
Phenotype	Asexual bloodstage;

Last modified: 11 November 2020, 17:12

*RMgm-4828

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene mutation
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 32340987
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	P. berghei ANKA 820cl1m1cl1 (RMgm-164)
Other information parent line	P. berghei ANKA 820cl1m1cl1 (RMgm-164) is a reference ANKA mutant line which expresses GFP under control of a male and RFP under control of a female gametocyte specific promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 19438517).
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The mutant parasite was generated by
Name PI/Researcher	Simwela NV, Waters AP
Name Group/Department	Institute of Infection, Immunity & Inflammation, Wellcome Centre for Integrative Parasitology
Name Institute	University of Glasgow
City	Glasgow
Country	UK
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Name of the mutant parasite
RMgm number	RMgm-4828
Principal name	G1808(V2721F)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Blood stages show different susceptibility to drugs (V2721F UBP mutation results in reduced artemisinin susceptibility)
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a mutated UBP1 (V2721F) Protein (function) Mutations in a ubiquitin hydrolase, UBP (HAUSP or USP7 close homologue), were previously identified to modulate susceptibility to ART and chloroquine (CQ) in the rodent infectious malaria parasite, Plasmodium chabaudi Phenotype Evidence is presented that V2721F UBP mutation results in reduced artemisinin susceptibility, while the V2752F mutation results in resistance to chloroquine and moderately impacts tolerance to artemisinins Additional information Other mutants

Mutated: Mutant parasite with a mutated gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0208800

Gene Model P. falciparum ortholog

PF3D7_0104300

Gene product

ubiquitin carboxyl-terminal hydrolase 1, putative

Gene product: Alternative name

UBP1

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Details of the genetic modification

Short description of the mutation

UBP1 mutated - V2721F

Inducable system used

Short description of the conditional mutagenesis

Not available

Additional remarks inducable system

Type of plasmid/construct

CRISPR/Cas9 construct: integration through double strand break repair

PlasmoGEM (Sanger) construct/vector used

Modified PlasmoGEM construct/vector used

Plasmid/construct map

Plasmid/construct sequence

Restriction sites to linearize plasmid

Selectable marker used to select the mutant parasite

unknown

Promoter of the selectable marker

unknown

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

See the paper for more details for the CRISPR/cas9 constructs and selection procedures

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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