2022.11.11 Information

EditForce achieved establishment of a high-throughput construction method for PPR protein gene and its application for splicing control in human cells

We established a technology of high-throughput construction of artificial PPR protein genes designed for the target RNA sequence in a joint study with Professor Takahiro Nakamura, Faculty of Agriculture, Kyushu University.
In addition, we have proven that we can apply PPR protein created using this method for exon skipping in human cells.
This study was published in the scientific journal Cells on November 8, 2022.

The novel PPR construction technology presented here improves the efficiency of PPR protein gene production and reduces the time required to select drug candidates for gene therapy using said technology.
Moreover, it is also expected to open up the possibility of gene therapy by exon skipping using PPR protein and contribute to the research and development of treatments for various genetic diseases.
We will push forward our R&D utilizing this technology to expand our drug discovery pipelines and create novel drugs.

Outline of Study

We have the technology to design PPR proteins that bind to specific sequences of target RNA. PPR proteins have a repeating PPR motif structure consisting of 35 amino acids. It has been challenging to create artificially designed PPR protein genes, and the steady and efficient production of a large number of PPR protein genes is time-consuming.
This study shows a way of optimizing of the sequence of PPR protein gene, a breakthrough technology that can easily connect multiple PPR motifs sequences without disrupting repeat sequences. We have also confirmed that PPR protein created utilizing this technology has a sequence-specific RNA binding activity.
By automation of this construction technology, we can produce large-scale PPR protein libraries quickly, enabling the efficient creation and discovery of PPR proteins that can be drug candidates.

In addition, we have proven that we can realize exon skipping of the designated target in human cells utilizing PPR protein, which can bind to RNA.
Exon skipping is a technique to artificially remove (skip) a portion in messenger RNA (exon) corresponding to a protein sequence.
Proteins are synthesized from messenger RNA, and various proteins are produced by complex changes in the combinations of exons that comprise the messenger RNA. Mutations within exons can prevent proteins from being appropriately synthesized, leading to the onset of particular diseases.
This research has opened up the possibility of new gene therapies by improving the abnormal composition of exons or removing exons.

Article information

Yusuke Yagi, Takamasa Teramoto, Shuji Kaieda, Takayoshi Imai, Tadamasa Sasaki, Maiko Yagi, Nana Maekawa and Takahiro Nakamura
Construction of A Versatile, Programmable RNA-binding Protein using Designer PPR Proteins and Its Application for Splicing Control in Mammalian Cells 2022, 11(22), 3529