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2017 年度 実績報告書

SINEUP ncRNAの解明: 期待される次代の分子生物工学技術ツール

研究課題

研究課題/領域番号 17F17775
研究機関国立研究開発法人理化学研究所

研究代表者

Carninci Piero  国立研究開発法人理化学研究所, ライフサイエンス技術基盤研究センター, 副センター長 (10333296)

研究分担者 RAHMAN MOHAMMED  国立研究開発法人理化学研究所, ライフサイエンス技術基盤研究センター, 外国人特別研究員
研究期間 (年度) 2017-11-10 – 2020-03-31
キーワードSINEUP / EEA elements / SINEB2 elements
研究実績の概要

Project 1: iPSCs reprogramming using CRISPRa EEA [EGA (embryo genome activation) enriched Alu-motif (EEA-motif)] motifs and SINEUPs - We have effectively designed SINEUPs against Yamanaka factors (OMKSL). We also checked basal expression level of OMKSL in human fibroblasts and iPSC. Moreover, we are optimizing different transfection protocol in human fibroblast and iPSC. Finally, we are designed and cloning necessary constructs.
Project 2: Dissecting physiological role of lncRNAs with SINEB2 elements - We have finalized the list of SINB2 containing lncRNAs (research plan; project2; a) that we want to dissect in details by analyzing different database analysis. We looked for their expression in different tissues and established cell lines and their involvement in different patho-/physiological functions. Moreover, we started with and got some initial data regarding research plan; project2; a) & b).

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

The main reason I feel like I lagged behind a little because of the major revision that we have made to our research proposal in order to accommodate recent advancement in the flied. But, we picked up the pace recently and hopefully we will match our research goal quickly.

今後の研究の推進方策

Future plan (project 1):
1) Transfection optimization (both fibroblast and iPSC), 2) Testing SINEUP against Yamanaka factors (OMKSL), 3) Testing CRISPRa against EEA elements, 4) Establish dedifferentiation of fibroblast into IPSC cells (in house)

Future plan (project 2):
1) Defining relationship between IncRNA/SINEB2 and sense protein coding genes by knockdown and overexpression experiments, 2) Optimizing protocols (RNA FISH, ChRIP-Seq/MS etc) to evaluate the mechanism of action of IncRNA/SINEB2.

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公開日: 2018-12-17  

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