Genomics-based improvement of Vero cells for virus growth

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14993
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental and hygienic pharmacy
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: Hanada Kentaro 国立感染症研究所, 細胞化学部, 部長 (30192701)
• Co-Investigator(Renkei-kenkyūsha): YAMAJI Toshiyuki 国立感染症研究所, 細胞化学部, 室長 (50332309) ; FUKASAWA Masayoshi 国立感染症研究所, 細胞化学部, 室長 (20291130) ; SAKUMA Chisato 国立感染症研究所, 細胞化学部, 研究員 (80782888)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 細胞基材 / ベロ細胞 / 宿主細胞 / ゲノム編集 / ウイルス / ワクチン / 感染症

Outline of Final Research Achievements

We recently examined the whole genome sequence of African green monkey kidney-derived Vero cells susceptible to various virus types. We herein prepared basic experimental conditions to produce new Vero cell variants that support better virus proliferation using genome editing technology. The following results were obtained. (1) Experimental conditions to isolate a Vero cell clone sustaining high susceptibility to Japanese encephalitis virus (JEV) were established. (2) The genome-wide screening of genes, the disruption of which confers Vero cells with resistance to Shiga-like toxin, was conducted. This screening hit many genes involved in the biosynthesis of the toxin receptor glycosphingolipid Gb3, which confirmed that the human CRISPR single-guide RNA library is applicable to Vero cells. (3) Among four Vero sublines, a parental Vero cell subline suitable for a somatic genetic approach was selected based on the stability of its karyotype and the sustainability of JEV susceptibility.

Free Research Field

感染細胞科学