Plant Chloroplast Genome Editing

Chloroplasts are semi-autonomous organelles with a genome separate from the nucleus and contain ribosomes for the translation and synthesis of their proteins. Chloroplast DNA is circular, generally between 120 and 170 kb in length, and has a molecular weight of 80 to 130 million Da. Most plant chloroplasts contain around 120 genes, most of which encode proteins required for photosynthesis and gene expression. With the sequencing of many chloroplast genomes in land plants and algae, chloroplast DNA has become a popular target for plant genetic research and crop improvement. As a result, chloroplast genome editing techniques have also received widespread attention.

Lifeasible has spent many years in the field of chloroplast genome engineering technology, overcoming the inability of many programmable genome editing tools to be applied to plant organelle DNA and developing our unique chloroplast genome editing solution. We guarantee to provide you with the most appropriate plant chloroplast genome editing services to help you explore chloroplast gene function, improve photosynthetic capacity, enhance food crops, and more.

What We Offer

Particle Bombardment

The particle bombardment is also known as high-velocity particle microprojector or gene gun bombardment. This method uses a power system to inject metal particles (gold or tungsten particles) with exogenous DNA into the plant chloroplast at a certain speed to achieve homologous recombination of genes. This gene editing technique must be applied to monocotyledons and is less efficient in transformation.

Golden Gate Cloning System

We can edit the chloroplast genome using the Golden Gate cloning system that scientists have developed.

• First, we construct the DdCBE Golden Gate target vector for expression in plant protoplasts according to your editing objectives.
• Next, in vitro transcription of the mRNA is performed.
• The protoplasts of the plant to be edited are isolated and transfected.
• And the protoplasts are cultured and the successful protoplasts are screened with screening antibiotics.
• The total DNA or RNA is extracted from the liquid medium and cultured cells in the transgenic healing tissue.
• Finally, libraries are prepared and targeted for deep sequencing.

CRISPR/Cas9 Editing System

Extensive research has led to the construction of a CRISPR-based targeted chloroplast genome editing system, nucleic acid constructs, vectors or vector combinations for targeted chloroplast genome editing, and a targeted chloroplast genome editing method. We can use this method to perform gene knockout or homologous recombination and targeted insertion of exogenous fragments easily and efficiently at your intended plant genomic locus.

Firstly, the Cas9 protein is brought into the chloroplast through the chloroplast signal peptide infA, thus achieving the knockout of genes on the chloroplast genome. Compared with the conventional particle bombardment technique, the method of the present invention not only reduces the difficulty of the operation and improves the efficiency and accuracy of targeted editing of chloroplast genes but also reduces the cost of operation. In addition, the present method can be effectively used for those plant species that cannot be transformed by particle bombardment.

Plant Species Covered by Our Service

Applications

• Production of edible protein
• Improvement of agronomic traits
• Production of vaccine proteins
• Production of antibody proteins
• Production of critical pharmaceutical proteins, etc.

Our Service Process

Lifeasible offers a customizable plant chloroplast genome editing service with excellent plant genetic engineers and extensive service experience. We sincerely look forward to working with you, and please feel free to contact us if you are in doubt.