RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4476

Malaria parasite	P. yoelii
Genotype
Tagged	Gene model (rodent): PY17X_0418900; Gene model (P.falciparum): PF3D7_0905300; Gene product: dynein heavy chain, putative (Dhc1) Name tag: GFP
Phenotype	Gametocyte/Gamete;

Last modified: 15 August 2018, 16:01

*RMgm-4476

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 28034675
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. yoelii
Parent strain/line	P. y. yoelii 17XNL
Name parent line/clone	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Liu, C; Yuan, J
Name Group/Department	State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Biology, School of Life
Name Institute	Xiamen University
City	Xiamen, Fujian
Country	China
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Name of the mutant parasite
RMgm number	RMgm-4476
Principal name	Dhc1::GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Not different from wild type
Gametocyte/Gamete	Dhc1::GFP expression in male gametocytes/gametes
Fertilization and ookinete	Not different from wild type
Oocyst	Not different from wild type
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a C-terminal GFP-tagged version of Dhc1. The mutant does not contain a drug-selectable marker For tagging Dhc1 the CRISPR/Cas9 method was used as described for mutants RMgm-1095 and RMgm-4071. Protein (function) A male gametocyte/gamete-specific dynein heavy chain protein Phenotype Dhc1::GFP expression in male gametocytes/gametes Additional information This mutant was used to cross with a mutant expressing mCherry in female gametocytes (RMgm-4475), resulting in selection of a mutant with GFP-expressing males (green) and mCherry expressing females (red) (RMgm-4477). All mutants are drug-selectable marker free. Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PY17X_0418900

Gene Model P. falciparum ortholog

PF3D7_0905300

Gene product

dynein heavy chain, putative

Gene product: Alternative name

Dhc1

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Details of the genetic modification

Name of the tag

GFP

Details of tagging

C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

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

hdhfr/yfcu

Promoter of the selectable marker

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

5-fluorocytosine (5-FC)

Additional remarks genetic modification

For tagging Dhc1 the CRISPR/Cas9 method was used as described for mutants RMgm-1095 and RMgm-4071.

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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