CRISPR enhances production of EPO used to treat anemia


Subscribe Us

By subscription, you consent to allow Ubigene Biosciences to store and process the information provided above to deliver the latest news, research spotlight, and promotions. You can unsubscribe from these communications at anytime.

Location:Home > About Us > Blogs >

CRISPR enhances production of EPO used to treat anemia

To many people, EPO rhymes with doping and cycling. But in fact, EPO is an important medical drug. This hormone works naturally in the body by stimulating red blood cell production. Patients suffering from anemia caused by for instance chronic kidney disease, AIDS or hematologic disorders can benefit immensely from EPO therapy. Furthermore, many cancer patients who are anemic from receiving chemotherapy are also in great need of EPO therapy. It is estimated that the market for EPO therapy is around 11 billion dollars annually.

Today, medical EPO is produced in mammalian cell lines using so-called recombinant DNA technology, where human genes are inserted to produce recombinant human EPO (rhEPO).

Recombinant human hormones such as EPO can be challenging to produce effectively, though, because they are quickly degraded by, amongst other things, the cells' own protein- and hormone-eating enzymes.

Using CRISPR technology, researchers from The Novo Nordisk Foundation Center for Biosustainability at Technical University of Denmark (DTU) have looked into how to reduce several cellular 'stressors' in order to improve production and product quality. This work has recently been published in the journal Metabolic Engineering.

The researchers found that by knocking out three genes involved in sugar group degradation and two involved in programmed cell death, they could improve production by 1.4-fold in Chinese Hamster Ovary (CHO) cells. Furthermore, the quality was greatly improved. The knockout cells contained around 40% of the most active EPO form, while the non-knockouts only contained around 2% of the most active EPO form (the highly sialylated EPO). The highly sialylated EPO form is important as it has the longest half-life. Only active EPO can treat conditions of anemia, so improving this feature is imperative in order to keep prices low and quality high.

How to avoid EPO degradation

The optimised cell line was actually designed to work in so-called fed-batch cultures, which is a way of feeding the cells continuously over the course of 10-14 days. Fed-batch is a commonly used method to make human hormones for various conditions and antibodies for cancer treatment.

But it is hard to produce EPO in fed-batch, because it is destroyed over time. Thus, EPO is nowadays produced in a 'one-feeding' manner (batch cultures), even though this method is less efficient. In order to do fed-batch cultures despite these obstacles, others have tried adding supplements to the mixture to avoid degradation. But with this new cell line, there is no need for workarounds, explains first author Tae Kwang Ha, Postdoc at The Novo Nordisk Foundation Center for Biosustainability:

"With our cell line, you don't have to add anything -- it just works."

These results have only been shown at small scale, so scale-up is needed to see if cells perform equally well in big production tanks. But this research verifies that knockout of specific genes can definitely improve productivity and product quality in single cells.

Story Source:

Materials provided by Technical University of Denmark. Original written by Anne Wärme Lykke and Anders Østerby Mønsted. Note: Content may be edited for style and length.

Journal Reference:

Tae Kwang Ha, Anders Holmgaard Hansen, Helene Faustrup Kildegaard, Gyun Min Lee. Knockout of sialidase and pro-apoptotic genes in Chinese hamster ovary cells enables the production of recombinant human erythropoietin in fed-batch cultures. Metabolic Engineering, 2020; 57: 182 DOI: 10.1016/j.ymben.2019.11.008

Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 1000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.

We developed CRISPR-U™ and CRISPR-B™ (based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, CRISPR-U™ and CRISPR-B™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. 

Genome Editing Platform
——Focusing on the Application of CRISPR-U™ and CRISPR-B™ Gene Editing Technology
1. Provides various types of gene-editing vectors for different species.2. Provides different virus packaging services, including lentiviruses, adenoviruses and adeno-associated viruses.3. Provides high-quality services for gene knockout, point mutation and knockin cell lines. 

Cell Biology Platform
——Focusing on primary cell
1. Provides over 400 types of primary cells.2. Provides culture strategies and related products for different cell types.3. Provides cell biology-related services such as cell isolation, extraction and validation.

Contact us