Method for mutation in organelle genome by CRISPR/CAS9

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K08453
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Parasitology (including sanitary zoology)
• Research Institution: Juntendo University
• Principal Investigator: HIRAI Makoto 順天堂大学, 医学部, 准教授 (50326849)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: マラリア原虫 / オルガネラ / ゲノム編集 / アトバコン / cytochrome b

Outline of Final Research Achievements

To deliver CAS9 protein into mitochondria, plasmids were constructed to replace nuclear localization signal with mitochondrial localization signal. Rodent malaria parasite, Plasmodium berghei was transfected with the plasmids, and localization of CAS9 protein in each transformant was investigated. As a result, no clear evidence for organelle localization of the protein was obtained. Beside, introduction of mitochondrial gene mutation by CRISPR/CAS9 (Mit) was attempted, however, desired mutation was not inserted in the genome. Taken together, these results suggest that delivery of CAS9 together with gRNA and donor DNA into organelle was critical point for successful modification of organelle genome. Further improvement should be challenged by considering these points.

Free Research Field

寄生虫学