Figure 19.

HEK293T cells exhibit neuronal properties upon induced expression of Arc. (A) Endogenous Arc expression was enhanced in HEK293T cells by targeting two single guide RNAs (sgRNAs) containing MS2 aptamers to the Arc promoter with the CRISPR/Cas9 Synergistic Activation Mediator system (see Methods for details). As a negative control, we used two sgRNAs targeting the promoter of the lac operon. Control cells (top) and Arc-induced cells (bottom) were stained for Arc (green) and DNA was labelled with DAPI (blue). About 90% of the cells expressed Arc. Scale bar is 30 µm. (B) Graph showing the top 20 differentially expressed genes upon the induction of endogenous Arc in HEK293T cells. RNA-Seq was used to compare the mRNA levels between the Arc-induced and control HEK293T cells. Neuronal genes are bolded and highlighted in red. (C,D) GO analysis of the differential expressed genes upon overexpression of Arc. The top 20 cellular components (C) and biological processes (D) are presented. Neuronal features were bolded and highlighted in red. Na⁺: sodium; V-gated: voltage-gated; Postsynapt: postsynaptic; Memb: membrane; Synapt: synaptic; Clath: clathrin; Ach: acetylcholine; Presynapt: presynaptic; Musc: muscle; Develop: development; Neg: negative; Reg: regulation; Prolif: proliferation; Skelet: skeletal; Contract: contraction and Mech: mechanical; Conv: conversion.