A Genome Editing Approach to Studying Pmp22 Enhancer Functionality

• DOI: 10.1096/fasebj.30.1_supplement.584.2
• Publication Date: 2023-11-26T16:52:35Z
• Authors: Harrison Pantera; John Moran; Camila Lopez‐Anido; John Svaren
• Keywords: Gene dosage

The function of the peripheral nervous system is dependent on myelination nerves by Schwann cells. This process tightly governed a network transcription factors during development. Myelin stability adversely affected in most common form hereditary neuropathy called Charcot‐Marie‐Tooth Disease. form, classified as CMT1a, caused 1.4 Mb duplication chromosome 17. includes abundantly expressed myelin gene Peripheral Protein 22 ( Pmp22 ), which associated with multiple disease states dosage‐dependent manner. Previous studies have shown that reducing expression rodent models CMT1A results amelioration phenotype found myelinating cells PNS. In recent work characterizing mechanisms regulating transcription, we identified cluster enhancers approximately 100 kb upstream TSS. was within smaller patients CMT1A‐like symptoms, where coding region itself not part duplication. possesses binding sites for are required cell development and myelination: Egr2 Sox10. our lab implicate these enhancers, but until now there has been an absence evidence functional importance achieving high endogenous levels expression. Using genome editing tools including CRISPR/Cas9 modified rat line expresses at near‐physiological levels, established clonal lines possessing deletions include factor‐binding sites. Preliminary data show observed decrease transcript ostensibly due to loss These first time requirement full alternate promoter usage. Support or Funding Information supported NIH grant R01 NS083841.