DNA methylation–independent long-term epigenetic silencing with dCRISPR/Cas9 fusion proteins

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

Abstract

The programmable CRISPR/Cas9 DNA nuclease is a versatile genome editing tool, but it requires the host cell DNA repair machinery to alter genomic sequences. This fact leads to unpredictable changes of the genome at the cut sites. Genome editing tools that can alter the genome without causing DNA double-strand breaks are therefore in high demand. Here, we show that expression of promoter-associated short guide (sg) RNAs together with dead Cas9 (dCas9) fused to a Krüppelassociated box domains (KRABd) in combination with the transcription repression domain of methyl CpG-binding protein 2 (MeCP2) can lead to persistent gene silencing in mouse embryonic stemcells and in human embryonic kidney (HEK) 293 cells. Surprisingly, this effect is achievable and even enhanced in DNA (cytosine-5)-methyltransferase 3A and 3B (Dnmt3A-/-, Dnmt3b-/-) depleted cells. Our results suggest that dCas9- KRABd-MeCP2 fusions are useful for long-term epigenetic gene silencing with utility in cell biology and potentially in therapeutical settings.

Details

Original languageEnglish
Article numbere202101321
Number of pages11
JournalLife science alliance
Volume5
Issue number6
Publication statusPublished - 14 Mar 2022
Peer-reviewedYes

External IDs

Scopus 85126724544
unpaywall 10.26508/lsa.202101321
PubMed 35288457
WOS 000771333100002
Mendeley 7e098785-c1b4-3070-afba-952fff9a5b17

Keywords

Keywords

  • Animals, CRISPR-Cas Systems/genetics, DNA Methylation/genetics, Epigenesis, Genetic/genetics, Gene Editing/methods, HEK293 Cells, Humans, Mice, RNA, Guide/genetics