Precise genomic correction by intercellular regulation of genome editing enzyme

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H05581
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Medical genome science
• Research Institution: Kyoto University
• Principal Investigator: Hotta Akitsu 京都大学, iPS細胞研究所, 特定拠点講師 (50578002)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ゲノム編集 / iPS細胞 / CRISPR / 相同組換え / 一塩基修復 / 一塩基変異

Outline of Final Research Achievements

About half of the genetic mutation causing human disease is a single nucleotide mutation. Deletion via NHEJ (nonhomologous end joining) can be relatively easily induced by using recent genomic editing techniques including CRISPR-Cas9, but knock-in using HDR (homologous recombination) efficiency was less than 1%, which was extremely inefficient. Therefore, we developed the CRONUS system in which CRISPR-Cas9 and guide-RNA were incorporated in the cells with a piggyBac transposon vector in advance, and Cas9 activity can be induced with two kinds of drugs. This made it possible to induce a desired single base modification at efficiencies as high as 20% to 30%, far exceeding the efficiency (<1%) so far.

Free Research Field

幹細胞遺伝子工学