Plant mitochondrial genome engineering: technology development and application to study fundamental aspects of mitochondrial gene expression

Introduction

PlaMitEng has the highly ambitious overall objective to make the mitochondrial genome of seed plants amenable to genetic engineering.

Project Overview

The project consists of two complementary work packages (WPs).

WP1: TALEN-GDM

In WP1, TALEN-GDM, a technology for plant mitochondrial genome editing recently developed by us will be used to systematically study plant mitochondrial gene expression in vivo.

Focus Areas

The focus will be on:

• Translation initiation
• Translational regulation in mitochondria

The mechanisms of which are currently completely unknown.

WP2: Direct Genetic Transformation

In WP2, PlaMitEng will develop a technology for direct genetic transformation of the plant mitochondrial genome.

Challenges

The mitochondrial genome is the only genome of the plant cell that, due to the lack of a mitochondrion-specific selection system, is not amenable to transformation and has remained recalcitrant to the insertion and expression of transgenes.

Strategies for Development

Taking advantage of:

1. Our recent construction of a mitochondrion-specific selectable marker gene
2. The generation of a dedicated recipient line for mitochondrial transformation
3. The design of a new transformation strategy (referred to as multivectorial transformation) that will enable the large-scale testing of expression elements for their suitability to drive the expression of mitochondrial transgenes in vivo

PlaMitEng will pursue the development of a robust technology for transformation of the plant mitochondrial genome.

Future Opportunities

The possibility to express transgenes from plant mitochondrial genomes will open up entirely new opportunities in both basic and applied research, and pave the way to harnessing the enormous potential of mitochondrial biotechnology and synthetic biology.