Single-Dose CRISPR/Cas9 Therapy Extends Lifespan of Mice with Hutchinson-Gilford Progeria Syndrome

Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare lethal genetic disorder characterized with symptoms reminiscent of accelerated aging. The major underlying genetic cause is a substitution mutation in the gene coding for lamin A, causing the production of a toxic isoform called progerin. Here we show the RNA sequencing analyses performed on primary mouse fibroblasts of different genotypes (in terms of Progeria; Cas9 background) treated with Cas9 guide RNAs targeting Lmna/Progerin. The mouse genetic background in (1) Pro/Pro;Cas9/+: homozygous for Progeria and heterozygous for transgenic Cas9 The mouse genetic background in (2) Pro/+; +/+: heterozygous for Progeria and no transgenic Cas9 (wild type) The mouse genetic background in (3) +/+;Cas9/+: no Progeria (wild type) and heterozygous for transgenic Cas9 Gene expression analyses of (1) Pro/Pro;Cas9/+ (2) Pro/+; +/+ (3) +/+;Cas9/+ primary mouse fibroblasts treated with a mock gRNA or Lmna target gRNA1, or Lmna target gRNA2. Each treatment was performed on fibroblast cells isolated from 2 different mice (2 biological replicates). The gRNAs were delivered into the cells by Lentiviral vectors that carried the mCherry gene as a reporter; no selection or sorting was performed post-treatment, however each sample was more than 80% mCherry-positive based on fluorescence microscopy. mock gRNA (a gRNA with no match on the genome) treatment was used as a negative control. Likewise, no transgenic Cas9 background (Pro/+; +/+) was used as another negative control.