Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of AP2-Z (see also RMgm-5032, RMgm-5034).

In addition, the mutant contains a gene encoding Cas9 from Streptococcus pyogenes, introduced into the c/d-ssu-rrna gene. This Cas9 nuclease does not cleave double stranded DNA in the absence of sgRNA and is thus suitable for generating the parasite which express it constitutively. It does not contain a drug-selectable marker which has been removed by negative selection.

The mutant does not contain a drug selectable marker

Protein (function)
In a previous study, an AP2 transcription factor-related gene, ap2r-1, was identified as a target gene of AP2-G and AP2-FG, and it was shown that its disruption affects ookinete development (RMgm-5032, RMgm-5034). Evidence is presented that AP2R-1 is an AP2 transcription factor that functions in zygotes; hence, it was renamed AP2-Z. 
AP2-Z has a putative AP2 domain that has not been identified by homology searches because of a non-canonical linker peptide between the second and third sheet. The domain is highly conserved in the Plasmodium species except for the linker peptide, which varies in amino acid sequence and length among the species.

The Cas9 nuclease does not cleave double stranded DNA in the absence of sgRNA and is thus suitable for generating the parasite which express it constitutively.

Phenotype
No GFP expression in female gametocytes. Strong fluorescence was observed in the nucleus of  AP2-Z::GFP zygotes.

From the paper: 'In our previous study (see RMgm-5034), we tagged AP2R-1 (AP2-Z) with mNeonGreen fluorescent protein (mNG)  using a conventional homologous recombination method (AP2R-1::mNG), and observed a fluorescent signal in the nucleus of female gametocytes, consistent with the previous observation that it is a target of AP2-G and AP2-FG. To evaluate whether ap2r-1 is actually transcribed by AP2-G and AP2-FG, we planned to perform a promoter assay on the endogenous ap2r-1. However, when we developed parasites expressing green fluorescent protein (GFP)-fused AP2R-1 (AP2R-1::GFP) by the CRISPR/Cas9 system using the Cas9-expressing parasite (PbCas9)  to perform the promoter assay, we observed no fluorescent signal in any blood-stage parasites, including female gametocytes.
In the homologous recombination method used previously, the 3’ untranslated region (UTR) of ap2r-1 was replaced by that of hsp70 upon tagging AP2R-1 with mNG. However, in this study, we inserted gfp at the 3’ end of ap2r-1 by CRISPR/Cas9 without replacing the original 3’ UTR. In female gametocytes of the Plasmodium species, a subset of genes is translationally repressed by the DOZI complex in a 5’ UTR- or 3’ UTR-dependent manner. Therefore, we assumed that ap2r-1 is translationally repressed by the DOZI complex in normal females  but translated in females of AP2R-1::mNG because of the replacement of 3’ UTR. Consistent with this assumption, strong fluorescence was observed in the nucleus of  AP2R-1::GFP zygotes.'

Other mutants