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DNA-seq (low-coverage WGS) for validation of CRISPR-Cas9-edited mESCs

ABSTRACT: DNA-seq (low-coverage WGS) for validation of CRISPR-Cas9-edited mESCs

PROVIDER: PRJEB61022 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR11176223.fastq.gz	Fastqsanger.gz
	ERR11176224.fastq.gz	Fastqsanger.gz
	ERR11176225.fastq.gz	Fastqsanger.gz
	ERR11176226.fastq.gz	Fastqsanger.gz
	ERR11176227.fastq.gz	Fastqsanger.gz

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DNA-seq (low-coverage WGS) for validation of CRISPR-Cas9-edited mESCs

Project description:Mouse embryonic stem cells (i.e., mESCs; line ESC 129-B13) were genetically modified using CRISPR-Cas9 to mutate the H3f3b locus, in order to carry homozygous lysine-to-alanine substitution of residues K9, K27 or K79. Two control mESC lines carrying knock-out of the H3f3a gene were used as background for the editing. Low-coverage whole-genome sequencing (~0.8/0.9X) was performed to assess the chromosomal integrity of the edited lines. Genomic DNA was extracted from each mutant and control mESC line and sonicated to obtain fragments of ~150-200bp on average. Fragmented DNA (~0.5-1 ug) was used for library preparation using the NEBNext Ultra II DNA library preparation kit (New England Biolabs). Sequencing was performed on a NextSeq500 platform in single-end mode (75bp reads).

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Transcriptomic profiling by RNA-Seq of MCF-7 cells edited by CRISPR/Cas9 in the FASN locus

Project description:We report a preliminary RNA-Seq data of MCF-7 cells edited in the FASN locus using CRISPR/Cas9. MCF-7 cells were edited by CRISPR/Cas9, screened by Surveyor assay, purified by limiting dilutions, validated by sanger sequencing, and characterized by diverse cellular analyses. Then, RNA from edited cells and non-edited controls were processed by TruSeq-RNA-Library-Prep-Kit and sequenced in a illumina equipment. Reads were aligned to hg38 in Galaxy using RNA-Star and transcripts were counted by FeatureCounts.

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