Development of a DNA memory that records cell-to-cell transfer of bacterial small regulatory RNA

2016 Fiscal Year Final Research Report

• Project/Area Number: 26670218
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bacteriology (including mycology)
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: Yamamoto Shouji 国立感染症研究所, 細菌第一部, 主任研究官 (80469957)
• Co-Investigator(Renkei-kenkyūsha): MITOBE JIRO 国立感染症研究所, 細菌第一部, 主任研究官 (40333364)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 細菌間コミュニケーション / 低分子RNA / RNA取り込み能 / RIVET

Outline of Final Research Achievements

This study was done to develop a DNA memory that records cell-to-cell transfer of bacterial small RNA (sRNA). This system consists of two distinct elements: one is a sensor of specific sRNA and the other is a memory of sensed information. The sensor is a translational fusion of the site-specific recombinase gene tnpR with a target gene of the sRNA while the memory is a DNA cassette containing a chloramphenicol (Cm) resistance gene and a levansucrase gene, which is flanked by TnpR-binding sites. In response to the presence of a specific sRNA, the expression of TnpR is activated. Synthesized TnpR in turn promotes excision of the DNA cassette, which causes sensitivity to Cm and confers resistance to sucrose. I constructed a series of sensors/memories for TfoR, the competence-activating sRNA in Vibrio cholerae, and evaluated performances. However, their low specificities and sensitivities were problematic. It is necessary to improve these issues for developing this type of memory device.

Free Research Field

細菌学