Identification of regulatory factors for genomic imprinting using siRNA library

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The aim of this study is to clarify regulatory mechanisms of the imprinting. (1) To distinguish imprinting disrupted cells from normal cells by expression of marker genes, Igf2r was targeted with YFP cassette on the maternal allele in F1ES cell. We are trying to introduce Igf2r-2A-E2Crimsoncassette into the paternal allele. The cells will be screened with siRNA library to identify regulatory factors for the imprinting. (2) non-coding RNA LIT] would recruit HDAC(s) to the promoter region of KvLQTl to deacetylate histone H3 and to make heterochromatin, leading to repress the KvLQTl gene on the paternal chromosome. (3) At mouse Commdl/U2af1-rsl imprinted domain, that interference with paternal Commdl transcription by the oppositely directed U2afl-rs1 transcription seemed to be a mechanism for the maternal predominantly expression of Commdl gene.