A CRISPR homing screen finds a chloroquine resistance transporter-like protein of the Plasmodium oocyst essential for mosquito transmission of malaria

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 2, 2024

SUMMARY Genetic screens with barcoded Plasmo GEM vectors have identified thousands of Plasmodium gene functions in haploid blood stages, gametocytes and liver stages. However, the formation diploid cells by fertilisation has hindered use genetic to investigate vector-parasite interactions during mosquito stages parasite. In this study, we developed a scalable system that uses targeting equipped CRISPR-mediated homing mechanism generate homozygous loss-of-function mutants reveal functionally life cycle system, knockout vector additionally expressing gRNA for its target is integrated into one parental alleles directs Cas9 intact allele after fertilisation, leading disruption. We find strategy 90% effective oocyst, resulting generation genotypes. A pilot screen reveals PBANKA_0916000 encodes chloroquine resistance transporter-like protein, CRTL, essential oocyst growth sporogony. The data point an unexpected importance transmission malaria poorly understood digestive vacuole contains hemozoin crystals. new screening provides method discover systematically at scale genes whose first are bloodmeal, enabling their potential as targets transmission-blocking interventions be assessed.

Language: Английский

A comprehensiveSchizosaccharomyces pombeatlas of physical transcription factor interactions with proteins and chromatin

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 20, 2024

SUMMARY Transcription factors (TFs) are key regulators of gene expression, yet many their targets and modes action remain unknown. In Schizosaccharomyces pombe , one-third TFs solely homology-predicted, with few experimentally validated. We created a comprehensive library 89 endogenously tagged S. TFs, mapping protein chromatin interactions using immunoprecipitation-mass spectrometry immunoprecipitation sequencing. Our study identified interactors for half the over quarter potentially forming stable complexes. discovered DNA binding sites most across 2,027 unique genomic regions, revealing motifs 38 uncovering complex regulatory network extensive TF cross- autoregulation. Characterization largest family revealed conserved sequence preferences but diverse patterns, repressive heterodimer, Ntu1/Ntu2, linked to perinuclear localization. TFexplorer webtool makes all data interactively accessible, offering new insights into mechanisms broad biological relevance. HIGHLIGHTS Comprehensive strain Experimentally determined atlas proteins web application interactive exploration interactomes Identification Nattou localization

Language: Английский