Development of a multiple knockout technology with CRISPR/Cas9 in rats

2014 Fiscal Year Final Research Report

• Project/Area Number: 25890011
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Laboratory animal science
• Research Institution: Kyoto University
• Principal Investigator: YOSHIMI Kazuto 京都大学, 医学(系)研究科(研究院), 研究員 (50709813)
• Project Period (FY): 2013-08-30 – 2015-03-31
• Keywords: ラット / ゲノム編集 / ノックアウト / ノックイン / CRISPR/Cas9

Outline of Final Research Achievements

Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system was applied as a newly and efficient genome editing tool in rats. We focused on the coat color related gene, Tyr, Asip and Kit genes. Microinjection of guide RNA (gRNA) and Cas9 mRNA into rat embryos produced a variety of insertion or deletion mutations and generated knockout (KO) rats by using the CRISPR/Cas9 system. Furthermore, combination of CRISPR/Cas9 and single-stranded oligodeoxyribonucleotides (ssODNs) enabled us to generate several types of targeted knock-in (KI) rats, such as SNP substitution, short DNA fragment integration, and large DNA fragment elimination. Therefore, CRISPR/Cas9 provides an efficient and robust genome engineering tool to generate multiple KO and KI rats.

Free Research Field

実験動物学