Inhibition of the PLOD2 gene by epigenetic editing
2020
University Medical Center Groningen, Groningen, Netherlands
Epigenetic editing is a new method for correcting pathological gene expressions. The present study investigated how to inhibit the PLOD2 gene, associated with fibrosis and cancer. For this purpose, the effect of DNA methyltransferase M.SssI and the KRAB repressor on PLOD2 expression in human fibroblasts and breast cancer cells was compared and analyzed using various genetic engineering methods. M.SssI induced de novo DNA methylation and altered the histone modifications contextually. Furthermore, a 50%-70% reduction in gene expression of PLOD2 was observed. The alignment of the KRAB repressor, which is routed to the PLOD2 promoter via zinc finger or CRISPR-dCas9-mediated targeting, caused the deposition of repressive histone modifications and resulted in almost complete suppression of PLOD2 expression. In both cases, the expression induced by the growth factor TGF-beta1 could be inhibited. Unstimulated PLOD2 expression, on the other hand, could only be detected by KRAB. In other kidney cancer and breast cancer cell lines, targeting temporarily expressed dCas9-KRAB induced ongoing PLOD2 repression. Based on the results, KRAB-induced heterochromatin enables effective gene compression without DNA methylation, which is simultaneously resistant to activations by TGF-beta1. PLOD2 repression by means of epigenetic editing can, for example, help to prevent tissue fibrosis or the formation of metastases and to develop individualized therapeutic approaches.
KRAB-induced heterochromatin effectively silences PLOD2 gene expression in somatic cells and is resilient to TGFβ1 activation
Marianne G. Rots
Added on: 08-22-2022
[1] https://www.mdpi.com/1422-0067/21/10/3634
