Gene delivery into the plant mitochondria via organelle-specific peptides

Dual Peptide-Based Gene Delivery System for the Efficient Transfection of Plant Callus Cells

“…7,8,14 Additionally, the carrier peptides composed of a polycation peptide and an organelle-targeting peptide achieved selective gene delivery to mitochondria and chloroplasts in several plant species 6,9,15 due to the high selectivity of the targeting peptides against these organelles. 34 In addition to the peptide-based gene delivery, nanoparticlebased approaches have emerged as alternatives to the traditional plant gene delivery methods. 35−39 For example, carbon nanotubes modified with polyethylene imine have been shown to enable pDNA delivery into the nucleus of several plant species without unexpected transgene integration, 37 whereas other carbon nanotubes functionalized with chitosan have achieved pDNA delivery to chloroplasts of different plant species.…”

“…To date, many gene carrier peptides have been developed and extensively tested in mammalian cells to achieve efficient nonviral gene therapy. , More recently, gene carrier peptides have been explored in plant cells and shown to deliver exogenous genes across the dual barrier consisting of the cell wall and plasma membrane. − The ability to delivery exogenous genes into plants can offer opportunities for enhancing crop yields and producing valuable pharmaceuticals or biofuels. − The existing plant gene delivery methods, such as the Agrobacterium method and biolistic-mediated delivery, are currently widely used but limited in terms of the sizes and types of deliverable DNA cargo, applicable plant types, and target organelles. , In contrast, peptide-mediated gene delivery has the potential to overcome these limitations. Polycation peptides are capable of condensing various sizes and types of DNA and forming peptide/DNA ionic complexes with tolerance toward nuclease-mediated degradation, − while cell-penetrating peptides (CPPs) have been shown to permeate the plasma membrane in a wide range of plant species and tissues. ,− To exploit these useful peptide properties, we designed gene carrier peptides obtained via the fusion of a polycation peptide to a CPP and found that the resulting carrier peptides successfully delivered plasmid and double-stranded DNA into plant cell nuclei. ,, Additionally, the carrier peptides composed of a polycation peptide and an organelle-targeting peptide achieved selective gene delivery to mitochondria and chloroplasts in several plant species ,, due to the high selectivity of the targeting peptides against these organelles …”

Rational Designs at the Forefront of Mitochondria-Targeted Gene Delivery: Recent Progress and Future Perspectives

“…Transfection of the constructed pDNA into the mitochondrial ATP6-mutated cell line critically improved ATP production, indicating the good functionality of the optimized MTS motif. MacMillan et al evaluated the computationally predicted MTS motifs for plant mitochondrial targeting using TargetP, which can predict the subcellular locations of proteins . TargetP prediction using selection criteria provided 748 candidates from 785 615 plant proteins.…”

A Synthetic Biology Workflow Reveals Variation in Processing and Solubility of Nitrogenase Proteins Targeted to Plant Mitochondria, and Differing Tolerance of Targeting Sequences in a Bacterial Nitrogenase Assay

“…The mitochondrial matrix has been shown to be a suitable location to express some of the most oxygen-sensitive Nif components in a functional form, including the Fe protein and NifB ( López-Torrejón et al., 2016 ; Burén et al., 2017a ; Burén et al., 2019 ). Although direct expression of nif genes from the mitochondrial genome is an attractive approach, it is currently technically difficult to directly introduce transgenes into the mitochondrial genome and recover stable transgenic plants ( Yoshizumi et al., 2018 ; MacMillan et al., 2019 ). In this study, we utilize the endogenous mitochondrial protein transport pathway for the expression of nuclear-encoded genes within the mitochondrial matrix ( Figure 1A ; reviewed by Ghifari et al., 2018 ).…”