DNA methylation–independent long-term epigenetic silencing with dCRISPR/Cas9 fusion proteins
Research output: Contribution to journal › Research article › Contributed › peer-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 language | English |
---|---|
Article number | e202101321 |
Number of pages | 11 |
Journal | Life science alliance |
Volume | 5 |
Issue number | 6 |
Publication status | Published - 14 Mar 2022 |
Peer-reviewed | Yes |
External IDs
Scopus | 85126724544 |
---|---|
unpaywall | 10.26508/lsa.202101321 |
PubMed | 35288457 |
WOS | 000771333100002 |
Mendeley | 7e098785-c1b4-3070-afba-952fff9a5b17 |
Keywords
ASJC Scopus subject areas
Keywords
- Animals, CRISPR-Cas Systems/genetics, DNA Methylation/genetics, Epigenesis, Genetic/genetics, Gene Editing/methods, HEK293 Cells, Humans, Mice, RNA, Guide/genetics