Cas9 Protein Post-translational Modifications (PTMs): A Potential Biomarker of Gene-editing
- Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)
The clustered regularly interspaced short palindromic repeats (CRISPR) arrays and the CRISPR associated (Cas) proteins comprise a prevalent prokaryotic and archaeal adaptive immune system. The CRISPR/Cas9 system has been coopted for and become the ubiquitous gene-editing system due to the simplicity of requiring minimally the CRISPR RNA components and Cas9 protein for specific DNA sequence alteration. CRISPR/Cas9 has been extensively used for gene-editing a wide range of species with human patient trails currently underway. However, unsanctioned genome editing is a national security and public health threat that can cause serious permanent illness and death as well as having the potential for very long-lasting effects over generations due to genetic inheritance of the gene-edit. While the Cas9 protein would appear as a highly specific indicator of exposure to gene-editing reagents, the bacterial origins of CRISPR/Cas9 creates a daunting problem for detection. Bacterial Cas9 would then generate false-positives for detecting gene-editing by conventional molecular biology techniques. Antibody-based assays for Cas9 would be unable to distinguish between Cas9 expressed in human cells for gene-editing and highly common unrelated Cas9 from bacterial infections. Posttranslational modifications of proteins are highly cell specific and hold the potential for discerning the cellular origins of a Cas9 protein and the differentiating between bacterial and gene-editing CRISPR/Cas9. The work described herein is in progress towards the identification of Cas9 post-translational modifications from bacterial and human cell expressed Cas9.
- Research Organization:
- Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)
- Sponsoring Organization:
- USDOE National Nuclear Security Administration (NNSA); USDOE Laboratory Directed Research and Development (LDRD) Program
- DOE Contract Number:
- AC04-94AL85000
- OSTI ID:
- 1571552
- Report Number(s):
- SAND--2019-12633R; 680482
- Country of Publication:
- United States
- Language:
- English
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