Single-cell genomics and genome editing of unculturable protist

2015 Fiscal Year Final Research Report

• Project/Area Number: 26660075
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied microbiology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Yuki Masahiro 国立研究開発法人理化学研究所, 環境資源科学研究センター, 特別研究員 (50572773)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: CRISPR / シロアリ / 共生原生生物

Outline of Final Research Achievements

The aim of this study was to perform genome editing of uncultivated symbiotic protists in the termite gut using the CRISPR/Cas system.
Cellobiohydrolase genes were detected in the symbiotic protist genome, and their target Cas9 sites were determined. We also calculated the codon usage of the protist genome, and t optimized the cas9 and GFP genes accordingly. The mixture of cas9 mRNA, gRNA and DNA for homologue recombination was injected into the termite gut. Furthermore, in order to remove contaminating sequences of symbiotic bacteria from the protist genome, we analyzed the genome of ectosymbionts by single-cell genomics.

Free Research Field

分子生物学