Fifth International Mammalian Synthetic Biology Workshop
Modulating Human Cas9-Specific Host Immune Response
Authors
Farzaneh Moghadam - Presenter, Arizona State University
Shayesteh Ferdosi, Arizona State University
Radwa Ewaisha, Arizona State University
Sri Krishna, Arizona State University
Samira Kiani, Arizona State University
Mo Ebrahimkhani, Mayo Clinic College of Medicine and Science
Karen Anderson, Arizona State University
Modulating Human Cas9-Specific Host Immune Response
As CRISPR technology is moving to the clinic, pre-existing immunity to SpCas9 in healthy individuals is a reasonable concern. We created modified Cas9 constructs by mutating bioinformatically defined human immunogenic epitopes and tested their function in comparison with wild-type Cas9. we examined variantsâ nuclease activity by targeting EMX-1 locus. Then, to investigate whether these variants retain the transcriptional modulation capacity, we targeted two different endogenous genes (TTN and MIAT).
Our data indicate that mutated Cas9 proteins recognize and bind their target DNA; leading to enhanced endogenous target gene expression or introduction of indels.