Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene tagging,
Gene tagging
|
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 38092766 |
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 2.34
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Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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The mutant parasite was generated by |
Name PI/Researcher | Hair M, Tewari R, Vaughan S |
Name Group/Department | Department of Biological and Medical Sciences |
Name Institute | Oxford Brookes University |
City | Oxford |
Country | UK |
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Name of the mutant parasite |
RMgm number | RMgm-5364 |
Principal name | SAS4-GFP/NDC80-mCherry |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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Phenotype |
Asexual blood stage | Not tested |
Gametocyte/Gamete | From the paper:
we analysed the positioning of the 8 NPs within the nuclear envelope. As the cell undergoes three rapid rounds of DNA replication, spindle microtubules nucleate from each NP and chromosome segregate as observed with kinetochore marker NDC80. The basal body marker SAS4-GFP is found associated with the kinetochore marker NDC80-Cherry even at 8 minutes after activation when nuclear content is octoploid as observed by live cell imaging. |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation
Parasites expressing GFP-tagged SAS4 (RMgm-5132) were crossed with parasites expressing mCherry-tagged NDC80 (RMgm-4844). After crossing gametocytes were analysed that express both SAS4-GFP and NDC80-mCherry (SAS4-GFP/NDC80-mCherry).
Protein (function)
SAS4 is a marker of the basal body (BB). SAS4/SAS6 is an ancestral core proteins involved in BB biogenesis.
NDC80 is a kinetochore marker. Chromosome attachment to spindle microtubules (MTs) is mediated by kinetochores, which are large multiprotein complexes assembled on centromeres located at the constriction point of sister chromatids
Phenotype
From the paper:
We analysed the positioning of the 8 NPs within the nuclear envelope. As the cell undergoes three rapid rounds of DNA replication, spindle microtubules nucleate from each NP and chromosome segregate as observed with kinetochore marker NDC80. The basal body marker SAS4-GFP is found associated with the kinetochore marker NDC80-Cherry even at 8 minutes after activation when nuclear content is octoploid as observed by live cell imaging.
A hallmark stage post-activation of gametocytes is the assembly of axonemes which are internally coiled around the nucleus and can clearly be seen at ~3-4 mins after activation using live cell imaging microscopy and basal body marker SAS-GFP with an axoneme marker Kinesin8B-mcherry.
Additional information
A hallmark stage post-activation of gametocytes is the assembly of axonemes which are internally coiled around the nucleus and can clearly be seen at ~3-4 mins after activation using live cell imaging microscopy and basal body marker SAS-GFP with an axoneme marker Kinesin8B-mcherry (see RMgm-5365)
From the Abstract:
Upon activation, the male P. berghei gametocyte undergoes three rounds of inter- nuclear mitosis and assembles eight basal bodies and axonemes around the nucleus prior to ex-flagellation, resulting in 8 flagellated male gametes in 12-15 minutes. In this study we used serial block face scanning electron microscopy (SBF-SEM) and cellular electron tomography (ssET) of P. berghei microgametocytes to examine the 3D architecture of key structures during male gamete formation. Our data has revealed an exquisite organisation of axonemes coiling around the nucleus in opposite directions forming a central ‘axoneme band’ in microgametocytes. Furthermore, we discovered that the nucleus of microgametes is tightly coiled around the axoneme in an exquisitely complex structure whose formation starts before microgamete emergence during ex-flagellation.
Other mutants |