Viral vectors, particularly AAV6, which are commonly utilized in gene editing, have been found to elevate p53 levels, thereby promoting cellular senescence in modified cells.

In mouse models, exposure to higher doses of AAV6 resulted in slower cell proliferation and increased senescence compared to control groups, which affected the balance of B and T cell production.

When targeting human hematopoietic stem and progenitor cells (HSPCs) with AAV6, researchers noted significant inflammation and elevated senescence markers, especially when focusing on the IL-2RG locus.

Researchers have investigated the causes of cellular senescence induced by CRISPR/Cas9 gene-editing technology, as detailed in a recent publication in Cell Reports Medicine.

The CRISPR/Cas9 process necessitates a double-strand break in DNA, followed by the insertion of new DNA and subsequent cellular repair, which activates a DNA damage response involving the p53 factor, ultimately leading to senescence.

While higher doses of AAV6 improved gene modification efficiency, they also heightened the likelihood of senescence in engineered cells, a phenomenon observed even in cells that did not express GFP.

To address the issue of senescence, researchers tested anakinra, an IL-1 antagonist, which successfully reduced senescence markers without compromising genetic engineering efficiency.

The use of anakinra not only decreased mutation rates but also minimized related genotoxicity, indicating its potential for clinical application in genetically engineered cells.

The findings suggest that pre-treatment with anakinra could become a standard procedure to mitigate unwanted side effects in gene editing for medical applications.

Additionally, targeting NF-κB with SC514 and p53 with GSE56 also resulted in reductions in senescence; however, these approaches increased mutation rates, raising concerns about potential genotoxicity.