RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1122000; Gene model (P.falciparum): PF3D7_0623100; Gene product: nuclear polyadenylated RNA-binding protein NAB2, putative (nab2)
PhenotypeNo phenotype has been described
Last modified: 26 October 2021, 17:20
Successful modificationThe 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 Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 34604117
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherNiikura M, Kobayashi F
Name Group/DepartmentDepartment of Environmental Science, School of Life and Environmental Science
Name InstituteAzabu University

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1122000
Gene Model P. falciparum ortholog PF3D7_0623100
Gene productnuclear polyadenylated RNA-binding protein NAB2, putative
Gene product: Alternative namenab2
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood-stage growth/multiplication.

Nuclear poly(A) binding protein 2 (NAB2), THO complex subunit 4 (THO4), nucleolar protein 3 (NPL3), G-strand binding protein 2 (GBP2) and serine/arginine-rich splicing factor 1 (SR1) are involved in nuclear mRNA export in malaria parasites.

NAB2 contains an N-terminal PWI domain and a C-terminal CCCH-type zinc finger motif, similar to yeast NAB2. Moreover, a nuclear localization signal (NLS), RX2–5PY NLS (PY-NLS), is present in NAB2.

To generated nab2-, tho4-, npl3- and sr1-deleted P. berghei ANKA, the gene-targeting vectors for nab2 (PBANKA_1122000), tho4 (PBANKA_1230500), npl3 (PBANKA_0506600) and sr1 (PBANKA_1232100) were prepared by PCR. Briefly, the 5’ and 3’ flanking regions of the open reading frame (ORF) of target genes were amplified by PCR. The PCR products were annealed to either side of the human dihydrofolate reductase (hdhfr)-expressing cassette and amplified by PCR using gene-specific primers The gene-targeting vectors were introduced into the ORFs of target genes by double-crossover homologous recombination
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