Genome Editing Plant Cells with Atmospheric Pressure Plasma

A team of researchers from the RIKEN Center for Sustainable Resource Science and partners attempted genome editing using the atmospheric-pressure plasma as protein introduction technique. Their findings are published in an open-access article in PlosOne journal.

Transgenic reporter plants were used as an experimental system to assess genome editing. The transgenic plants carried the reporter genes L-(I-SceI)-UC and sGFP-waxy-HPT. The success of genome editing was measured through the chemiluminescent signal from the refunctionalization of the luciferase gene that proceeded after genome editing. In the same way, the sGFP-waxy-HPT system led to hygromycin resistance caused by hygromycin phosphotransferase during genome editing.

CRISPR-Cas9 ribonucleoproteins, which target the reporter genes, were introduced into rice calli or tobacco leaves after treatment. Then the rice calli were placed on a suitable medium, leading to a unique luminescence signal. Meanwhile, the tobacco cells carrying the sGFP-waxy-HPT gene showed hygromycin resistance. After recurring cultivation of the treated tobacco leaf pieces on the regeneration medium, the calli grew leaf pieces. Hygromycin-resistant green callus was eventually harvested, and a genome-edited sequence in the tobacco reporter gene was confirmed.

Find more details in the study from PlosOne.