A biotech company pioneering transposases for cell line engineering claims to have widened the applications of these tools.
Oren Beske, PhD, gave a talk at BioProcess International, about how ATUM is pioneering the use of multiple transposase enzymes to engineer Chinese Hamster Ovary (CHO) cells for biologics manufacture.
“The industry is filled with many hard problems and people looking for unique ways to solve them and, because we have many of these enzymes, you don’t need to limit yourself to one category of genetic modification or insertion,” explains Beske.
“Whether you’ve got a toxic protein you want to express, or whether you want to knock down or overexpress something, you can be creative and think outside the box!”
According to Beske, ATUM led the charge in first harnessing transposase enzymes as a tool for creating cell lines to express proteinaceous drug.
By 2023, he says, single transposase enzymes were the industry standard for engineering cell lines to express a vaccine or antibody.
However, to make multiple genetic changes to a cell line, companies need to turn to other engineering methods, such as CRISPR. “Because, if you only have one enzyme, you run the risk that using it to do multiple things in sequence could lead to the initial transposon insertions becoming unstable,” he says.
To get around this problem, he says that ATUM has pioneered the use of multiple transposase enzymes. These are orthogonal, meaning that, if one enzyme is used to insert DNA into the genome, the second enzyme has no risk of affecting that change.
“By having multiple enzymes that do not recognize each other’s cognate transposons [pieces of ‘jumping DNA’ inserted into the genome by a transposase], we can now iteratively engineer cells to change their genetics and physiology,” he says.
In his talk, Beske presented a case study where ATUM used three enzymes to make three changes inside a CHO cell, which—in turn—created a cell line for expressing a uniquely glycosylated antibody. “The takeaway is the ability to think creatively,” he says.
Going forward, he says the company is developing a protein expression system based on multiple transposases, which can be used by its customers to create cell lines that can be used from high throughput in early-stage drug discovery to commercial manufacturing.
“As biologics become increasingly complex, we’ve always been faced with the challenge of translation from academic research to manufacturing,” he says.
“Making these complex proteins and unique biological entities is getting increasingly complex, and—what this means—is that a generalized CHO host may not be the best way to do that.”
He adds, “Manufacturing platforms will need to be more bespoke in the future.”