GSM8525891: High IRES,Cap Replicate 1; synthetic construct; OTHER
For RNA-Seq, full-length cDNA was prepared using the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing  (Takara Bio USA, Inc.) For CRISPRi, genomic DNA was isolated using NucleoSpin Blood Maxi kit (Macherey-Nagel, 740950). RNA-Seq libraries prepared using the Nextera XT DNA library preparation kit (Illumina) CRISPRi sgRNA target regions were PCR amplified  with Illumina adaptors and indexes using Ultra II Q5 (NEB Biolabs, M0544X) and products were purified  with DNA Clean & Concentrator-100 (Zymo Research, D4030). sgRNA target regions (260-280bp) were further enriched by gel extraction (Qiagen, 28704) after running agarose gel Smart-seq v4