Project description:To inverstigate off-target effects of CRISPR-iPAS, we transfected HEK293TAPA reporter cells with dPguCas13b and gUSE (showed most effective APA-interference) or non-targeting gRNA (gNT) respectively. 3'end-seq and RNA-seq were performed to investigate if the global change on the endogenous PAS usage and gene expression caused by CRISPR-iPAS. Comparing with gNT, dPguCas13b with gUSE showed high specificity.
Project description:We used a new technology, named Detect-seq, to perform genome sequencing on transfected HEK293T cells to see DdCBEs' off-target mutations. Besides, targeted-amplicon sequencing and ATAC-seq data were applied to validate the results of Detect-seq.
Project description:We used a new technology, named Detect-seq, to perform genome sequencing on transfected HEK293T cells to see DdCBEs' off-target mutations. Besides, targeted-amplicon sequencing, ATAC-seq and in situ ChIP-seq data were applied to validate the results of Detect-seq.
Project description:RNA-seq experiment to study the role of Pat1b in HEK293T cells. HEK293T cells were treated twice with siRNA with Lipofectamine 2000 then harvested after 48hours. Total RNA was extracted with TriReagent. Ribo-Zero TruSeq stranded mRNA libraries were prepared for each sample and sequenced on Illumina NextSeq 500 Sequencing System providing around 100 million reads per sample (around 75 bp paired-end reads).