Genome-wide Programmable Transcriptional Memory by CRISPR-based Epigenome Editing
By James K. Nuñez, Jin Chen, Greg C. Pommier, J. Cogan, J. Replogle, C. Adriaens, Gokul N. Ramadoss, Quanming Shi, King L. Hung, Avi J. Samelson and 10 more. Cell, 2021
604 citations recorded by the source corpus.
SUMMARY A general approach for heritably altering gene expression has the potential to enable many discovery and therapeutic efforts. Here, we present CRISPRoff—a programmable epigenetic memory writer consisting of a single dead Cas9 fusion protein that establishes DNA methylation and repressive histone modifications. Transient CRISPRoff expression initiates highly specific DNA methylation and gene repression that is maintained through cell division and differentiation of stem cells to neurons. Pairing CRISPRoff with genome-wide screens and analysis of chromatin marks establishes rules for heritable gene silencing. We identify sgRNAs capable of silencing the large majority of genes including those lacking canonical CpG islands (CGIs) and reveal a wide targeting window extending beyond annotated CGIs. The broad ability of CRISPRoff to initiate heritable gene silencing even outside of CGIs expands the canonical model of methylation-based silencing and enables diverse applications including genome-wide screens, multiplexed cell engineering, enhancer silencing, and mechanistic exploration of epigenetic inheritance.
Subfields: Molecular Biology
View Paper: https://www.semanticscholar.org/paper/7ab01c80a6442103e1226c286af71577733cd31a
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Citations in the corpus
- Mapping information-rich genotype-phenotype landscapes with genome-scale Perturb-seq (2021, methodology): “For bothK562andRPE1growth screens,DNA libraries of the initial and final sampleswereprepared for deep sequencingby genomic DNA isolation and PCR amplification of dual-sgRNA amplicons as described previously (Nuñez et al., 2021; Replogle et al., 2020).”