The Next Generation
Genome Editing Technology

RIDE of GERNA Biotech

4th tech.

No Cas system

More Safety

Lower error rate, off-target

wide coverage

No PAM sequence dependency

Achieve Your Goals
with Strategy

GERNA Biotech Co., Ltd. researches and develops gene therapy agents utilizing self-developed next-generation gene editing technology.

What we do

Gene editing technology has developed from the first generation of ZFN to the second generation of TALEN and the third generation of CRISPR-Cas9.

CRISPR-Cas9 was seen as a technique that could eliminate the fundamental problems of genetic diseases.

However, side effects in the double-strand breaks and repair process of CRISPR-Cas9 (DSB issue), off-target issues caused by short guide-RNA, and limitations in the applicable range because of dependence on PAM sequence are major obstacles to the development of various therapeutic agents.

Many studies are trying to improve the function of Cas and excellent techniques such as prime-editing have been developed. However, the problems of side effects in the DNA break and repair process, off-target issues from short guide-RNA and limitations because of PAM sequence dependence still exist.

1st POC of RIDE

The simple POC in human cells

The gene editing technique developed by Gerna Biotech does not use Cas and can be considered a fourth-generation gene editing technology.

It is a new method utilizing highly designed RNA and intracellular DNA repair mechanisms.

The non-Cas technique is free from DSB issues because there is no DNA break or repair process. In addition, off-target issues occur less because guide-RNA longer than that of the conventional methods is used. Since this technique is not dependent on PAM sequence, it can be applied to mutations that cause numerically 75% of all genetic diseases.

Gerna Biotech’s RNA-instructed DNA editing technology, abbreviated as RIDE technology, is a next-generation and fourth-generation gene editing technology.

2nd Approach We Follow

Project Workflow

Gerna Biotech Co., Ltd. demonstrated its RIDE technology through simple experiments.

1. Single nucleotide polymorphism mending

– Clone the GFP gene with intentionally applied SNP into the vector.

– Transfect the human cell line (HEK293T) using the vector containing broken GFP.

– The cells do not express fluorescence because GFP has been broken.

– Transfect the cell using the vector that can mend mutation in GFP and express the highly designed RIDE.

– RIDE is expressed in the cell and SNP in GFP is mended through intracellular repair mechanisms.

– The cell with normal GFP expresses GFP, exhibiting fluorescence.

2. Multiple mutation mending

– Clone the GFP gene with intentionally applied two SNPs of different types into the vector.

– Transfect the human cell line (HEK293T) using the vector containing broken GFP.

– The cells do not express fluorescence because GFP has been broken.

– Transfect the cells using the vector that can mend multiple mutations in GFP and express the highly designed RIDE.

– RIDE is expressed in the cell and multiple mutations in GFP are mended through intracellular repair mechanisms.

– The cell with normal GFP expresses GFP, exhibiting fluorescence.

3. Deletion mending

– Clone the GFP gene with several types of the intentional deletion into the vector.

– Transfect the human cell line (HEK293T) using the vector containing broken GFP.

– The cells do not express fluorescence because GFP has been broken.

– Transfect the cells using the vector that can mend deletion in GFP and express the highly designed RIDE.

– RIDE is expressed in the cell and deletion in GFP is mended through intracellular repair mechanisms.

– The cell with normal GFP expresses GFP, exhibiting fluorescence.

3rd Approach We Follow

POC in human genome

GERNA Biotech Co., Ltd. directly edited human genomes in human cell lines and cancer cells using RIDE.

1. Generating gene mutations related to rare genetic diseases

– Find well-known rare genetic disease-causing genes in the human cell line (HEK293T).

– We targeted well-known mutations related to cystic fibrosis (CFTR_G551D), AATD (SERPINA1_E366K), genetic hearing loss (GJB2_R143W), hereditary encephalopathy (RNH2B_A177T) and Hunter’s syndrome (IDS_R468Q).

– Design the RIDE to generate mutations at the well-known mutation site of each gene.

– Transfect the cells using the highly designed RIDE in the form of oligo-nucleotides.

– Check the mutagenesis in the gene based on the ddPCR result.

2. Editing cancer-related genes

– Select well-known cancer-related genes in the human cell line (HEK293T).

– Design the RIDE that generates mutations at the well-known mutation site of the genes: p53, KRAS_G12S, KRAS_G13D, PIK3CA_H1047R

– Transfect the cells using the highly designed RIDE in the form of oligo-nucleotides.

– Check the mutagenesis in the gene based on the ddPCR result.

3. Editing genes in cancer cells

– Select well-known cancer-related genes in the cancer cell line (Hela).

– Design the RIDE that generates mutations at the well-known mutation site of the genes: GRIK2_A490V, ZNF292_H1542R

– Transfect the cells using the highly designed RIDE in the form of oligo-nucleotides.

– Check the mutagenesis in the gene based on the ddPCR result.

4th Approach We Follow

Project Workflow

Gerna Biotech Co., Ltd. is researching and developing gene therapy agents through RIDE technology. The pipeline below is for ongoing in vivo experiments.

1. Gene therapy agents that eliminate mutations in p53, a cancer suppressor gene.

Mutations in the tumor suppressor gene p53 gradually increase because of aging and stress. If there are lethal mutations, cancer may occur. In the majority of patients, it has been found that many genetic mutations occur randomly. With the CRISPR-Cas method, drugs for each mutation should be developed. However, the RIDE method can provide immediate treatment if only the sequence to be repaired is known.

2. Longevity gene SIRT6

Centenarians have mutations in SIRT6 in common. Modification of the SIRT6 gene in general people will benefit all humankind. Since the site is at about 35mer level, editing the site with current CRISPR-Cas9 is difficult. Theoretically, it is possible to remove the entire site using the CRISPR-Cas system, followed by inserting DNA-fragments. However, there is a limitation in using the CRISPR-Cas system because the probability of mutagenesis in the DSB repair process is too high and it is challenging to predict side effects in genes that play various roles like TASL.

Since RIDE is a technology that can edit multiple mutations without the DNA break process, it is possible to develop corresponding gene therapy agents.

3. Obesity gene TASL

It is known that deletion in TASL can cause obesity. Theoretically, it is possible to cut the DNAs in part with deletion using the CRISPR-Cas system followed by inserting DNA-fragments. However, there is a limitation in using the CRISPR-Cas system because the probability of mutagenesis in the DSB repair process is too high and it is challenging to predict side effects in genes that play various roles like TASL. Since RIDE is a technology that can repair gene deletion without the DNA break process, it is possible to develop corresponding gene therapy agents.

Gerna Biotech Co., Ltd. aims to provide comfortable lives for humankind by mending DNA mutations, the root cause of disease, with RIDE, our next-generation gene editing technology.

Achieve Your Goals
with Strategy

The goal and strategy of GERNA Biotech

In vitro
In vivo
Patent