An efficient lentiviral CRISPRi approach to silence genes in primary human monocytes [article]

Bingxu Liu, Nir Hacohen, Darrell Irvine, Matteo Gentili, David Lieb, Thomas Eisenhaure
2020 bioRxiv   pre-print
Human primary monocytes are critical in controlling immune responses and tissue homeostasis. However, to identify and study molecular components that underlie the function of these cells, there remains a need to develop methods to perturb genes in these cells. Here we report lentiviral-based delivery of dCas9-KRAB and guide RNAs to efficiently silent target genes, such as CD45 and CD209. We further show that sgRNAs against TICAM1 dampen proinflammatory cytokine and interferon expression in
more » ... expression in response to lipopolysaccharide. When delivered by lentivirus, sgRNAs are incorporated into the genome, thus enabling pooled screening to perturb and identify coding and non-coding elements that contribute to the functions of primary human monocytes.
doi:10.1101/2020.12.23.424242 fatcat:eqtmtoumpbdb3fpboiqp63kxhu