Project description:wild-type and safa-/- THP-1 cells, or Flag-SAFA and Flag-Del-RGG stable expressed SAFA-/- cells were infected with VSV for 24 hours, samples were collected and send for RNA-seq.
Project description:Genome wide RNA-Seq screen was did to detect gene expression of HSV-1 and Hela . We constructed a HeLa cell line that stably expresses the plasmids of SAFA or SAFA-ADAR1 catalytic domain(SAFA-ADAR1cd) or SAFA-ADAR1 catalytic domain E488Q(SAFA-ADAR1cd E488Q) . The RNAs were harvested followed by RNA sequencing to identify the edited RNA.
Project description:Flag-SAFA or Flag-Del-RGG mutant stablely expressed THP-1 cells were infected with VSV for 6 hours, samples were collected and send for ATAC-seq.
Project description:We report long-read sequencing results from the sox32 allele targeted with a multi-guide Cas9 RNP strategy. Embryos were cell-injected with Cas9 RNP programmed with 4 guides simultaneously targeting different portions of the sox32 gene. Genomic DNA was extracted from each embryo and the sox32 coding sequence and intron were amplified and sequenced. Six individual embryos were evaluated. We found that the majority of sequences were unique and most involved mutations confined to the target sites rather than larger lesions spanning two or more target sites. We also found that alleles accumulated mutations over several DNA replication cycles, which may in part explain the diversity of alleles that were generated. This study is the first to use high-throughput sequencing to evaluate of an allele of this size mutated by a multi-guide Cas9 RNP strategy and provides insights into the dynamics of CRISPR-Cas9 RNP gene editing in vivo.