Trisomic rescue induction by allele-specific chromosome breakage using genome editing technique

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K15242
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Human genetics
• Research Institution: Mie University
• Principal Investigator: Hara Mari 三重大学, 医学部, 教務職員 (30176383)
• Co-Investigator(Kenkyū-buntansha): 脇田 幸子 三重大学, 医学部, 技術員 (20782981)
• Co-Investigator(Renkei-kenkyūsha): Hashizume Ryotaro 三重大学, 大学院医学系研究科, 助教 (50456662) ; Ichishi Masako 三重大学, 医学部, 技術専門員 (80632372) ; Miyagawa yoshitaka 日本医科大学, 大学院医学系研究科, 講師 (90415604) ; Inui masafumi 国立研究開発法人国立成育医療センター, システム発生/再生医学研究部, 室長 (20643498)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: トリソミックレスキュー / ハプロタイプフェイジング / 核型正常化 / CRISPR/Cas9 / ゲノム編集 / Cre-loxP

Outline of Final Research Achievements

We have successfully established three induced disomy 21 iPS cell lines by genome editing technology from the original trisomy 21 iPS cells derived from a single individual with Down's syndrome. These induced three types of cells have different combinations of 21 chromosomes. The karyotypes of the induced disomy cells have been confirmed by means of chromosome spreading G-banding, short tandem repeat analysis, multiplex ligation-dependent probe amplification, and fluorescent in situ hybridization. Furthermore, we also classified them based upon the origin of deleted chromosome 21 by STR analysis. Following chromosome phasing by comparison of sequencing data with the 3 cell lines, we have subsequently succeeded in constructing a CRISPR/Cas 9 system for allele specific cleavage in multiple sites.

Free Research Field

病理学・発生学