Aldevron Breakthrough Blog

Collaboration Helps Drive Advancement

September 27, 2023 / by Patrick Paez, Ph.D.

Making use of ultra-low endotoxin Cas9 from Aldevron

The latest research article from the Innovative Genomics Institute (IGI) at the University of California, Berkeley, highlights the collaboration between Aldevron with the laboratories of Nobel Laureate Dr. Jennifer Doudna and Dr. David Savage. Published in Molecular Therapy-Nucleic Acids, “Genome editing in the mouse brain with minimally immunogenic Cas9 RNPs,” shows how Aldevron generated ultra-low endotoxin Cas9 for IGI to enable a novel in vivo CRISPR delivery system, targeted directly into the brain for editing.

The study compared adeno-associated virus 9 (AAV9), the current “gold standard” for central nervous system gene editing, to IGI’s direct injection of the mouse striatum with cell-penetrating Cas9-RNPs termed convection-enhanced delivery (CED). Alternative delivery systems of AAV for CNS disorders remain a theme well entrenched within the field due to:

Authors of these types of reports often tell stories that required thousands of hours of development and were years in the making. In this publication, readers are guided through the journey using several graphics, summarized as:

Figure 1: Authors show in vivo proof of concept that delivering Cas9 RNPs via direct injection into the brain's striatum can edit neurons like AAV9-mediated delivery. They test two different injection catheters to optimize CED.

Figure 2: Discovered measurable levels of immune responses from both treatment modalities, prompting them to seek a lower endotoxin version of their Cas9.

Figure 3: Cas9 RNPs manufactured using an optimized low endotoxin Cas9 manufactured by Aldevron* reduced local immune response and adaptive immune responses.

The authors demonstrated a viable alternative in obtaining ultra-low endotoxin nuclease with the delivery of high-grade RNPs to meet the FDA required thresholds and an alternative to viral vector delivery (Supplemental figure 12A). This study shows the work at each step as obstacles were encountered in developing a potentially safe and translational method of delivering Cas9 RNPs via CED. This endeavor to reduce the endotoxin of Cas9 to meet FDA standards for intrathecal injection was a challenge Aldevron was ready to take on.

“Advancing every day means helping amazing researchers. The timeline for developing and manufacturing their Cas9 presented a challenge, but we operate with a sense of urgency. We don’t like to use the word impossible with our client's timelines,” said Tom Foti, VP of the Protein Business Unit, when Aldevron was approached for assistance with the project.

And as Athen Kimberlin, a Group Leader in Aldevron’s Protein Business Unit, points out, the customized protein with a low-endotoxin process was delivered within six weeks of the initial call. “There was definitely a buzz,” he said.

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*Aldevron provides RNPs only to customers who are duly licensed, including to make and have made RNPs, for their intended use.


Patrick Paez, Ph.D.

Patrick Paez, Ph.D.

Patrick Paez serves as a technical marketing manager for Aldevron’s marketing team. He earned a bachelor's degree in Pre-Medicine and Psychology, and a Ph.D. in Immunology at Virginia Commonwealth University (VCU) in Richmond, VA. His Ph.D. training at VCU's Medical College of Virginia campus was in a translational science lab at the Massey Cancer Center, where he focused on developing immunotherapeutic interventions in the field of immuno-oncology for phase 1 clinical trials.