Unknown,Transcriptomics,Genomics,Proteomics

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4C-seq to show the effects of insertion of STITCH into NEUROG2-65kb positions on the chromatin conformation in neural progenitor cells differentiated from human iPSCs.

ABSTRACT: We reconstituted arrays of CTCF binding sites (L1, L2, L3, L4, R1, R2, and R3) and devised a synthetic topological insulator with tetO for chromatin-engineering (STITCH). By coupling STITCH with tetR linked to the KRAB domain to induce heterochromatin and disable the insulation, we developed a drug-inducible system to control gene activation by enhancers. We inserted STITCH into the 65-kb downstream of NEUROG2 in human iPSCs and integrated a transgene consisting of tetR-KRAB followed by DNA encoding the 2A peptide and the puromycin resistant gene with piggyBac transposition into the genome in the STITCH/NEUROG2 clone (NEUROG2/KRAB). We performed 4C-seq (Circular chromatin conformation capture assay followed by deep-sequencing) from the viewpoint designed at R3 of the inserted STITCH in the differentiated neural progenitor cells with and without DOX.

INSTRUMENT(S): Illumina iSeq 100

ORGANISM(S): Homo sapiens

SUBMITTER: Taro Tsujimura

PROVIDER: E-MTAB-8957 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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4C-seq to show the effects of insertion of STITCH into NEUROG2-65kb positions on the chromatin conformation in neural progenitor cells differentiated from human iPSCs.

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