GSM3854794: L2 p5; Toxoplasma gondii; OTHER
DNA was isolated using the Qiagen Blood and Tissue kit, following the protocol for blood cells. Integrated gRNAs were amplified, barcoded, pooled by QuBit and sequenced on an Illumina MiSeq. We assembled two libraries, each targeting ~100 potential nucleic acid-binding proteins, with 5 gRNAs per gene. Library 1 (L1) largely consists of genes identified as differentially regulated in our preliminary RNA-seq experiment. Library 2 (L2) contains genes with DNA-binding domains commonly found in transcription factors, such as zinc finger and Myb-like domains. Across both libraries, all 67 members of the ApiAP2 transcription factor family are targeted, along with 151 putative nucleic acid-binding proteins. As controls, each library additionally contains 10 genes known to be essential, 10 genes known to be dispensable, 10 non-cutting gRNAs, and 5 gRNAs against the mNG reporter itself. Oligonucleotide pools were ordered from agilent, and assembled by Gibson assembly.
GEO Accession
GSM3854794