The Plasmodium falciparum CCCH zinc finger protein ZNF4 (PF3D7_1134600) plays an important role in gametocyte exflagellation through the regulation of male gametocyte enriched genes.

Purpose: To carryout comparative transcriptomics between the ZNF4-K0 and the wild type in the ring and mature gametocyte stage of Plasmodium falciparum Methods: Total RNA was isolated from ring stage and percoll- enriched mature (stage V) gametocytes from the ZNF4-KO and NF54 wildtype using the Trizol reagent (Invitrogen) according to the manufacturer’s protocol. . RNA sequencing was performed at the Genomic Facility of the University Clinic at the RWTH University Aachen, Germany. Briefly, the quality of the isolated total RNA samples from ring stage and mature gametocytes of the ZNF4-KO and the wildtype were evaluated by Tapestation 4200 (Agilent). Libraries were generated with TruSeq Stranded mRNA Library Preparation kit (Illumina) from high quality total RNA samples according to the manufacturer´s protocol. The generated libraries, which pass the QC check on Tapestation 4200 (Agilent) were sequenced on a NextSeq 500 (Illumina) with High output Kit v2.5 (150 cycles) for paired-end sequencing according to standard procedure provided by Illumina. Results:Comparative transcriptomics between wildtype and ZNF4-KO in ring and mature gametocyte stages show only 66 genes de-regulated by greater than 2-fold in the ZNF4-KO ring stage while about 473 (274- upregulated and 199 down-regulated) were deregulated in mature gametocytes. Conclusions: Our combine data therefore reveal that ZNF4 plays an important role in male gametocyte exflagellation through the regulation of male gametocyte genes Total RNA was isolated from ring stage and percoll- enriched mature (stage V) gametocytes from the Plasmodium faciparum ZNF4-KO and NF54 wildtype using the Trizol reagent (Invitrogen) according to the manufacturer’s protocol. Quality of RNA samples were assessed using a ND-1000 (NanoDrop Technologies, Thermo Scientific) and by agarose gel electrophoresis.RNA sequencing was performed at the Genomic Facility of the University Clinic at the RWTH University Aachen, Germany.