Figure 1. CRISPR-activation of collagens increases lifespan and heat stress resilience:A) Schematic illustration of bacteria feeding expressing
col-120 via promoter-specific sgRNA using the L4440-Biobrick-sgRNA plasmid to dCas9::VP64 transgenic C. elegans. B, D) Relative fold change in mRNA expression of
col-120 gene in young adult (YA) and day three adults, respectively, when fed sgRNA
col-120 or scr to eggs (B) or L4 (D) animals. qRT-PCR experiment was performed in three or more biological replicates. Welch's two-tailed t-test was used for significance analysis. C, E-J) Survival curves showing extension in lifespan in animals with a dCas9-activated expression of
col-120 (C, E-G) and
col-10 (H-J) compared to scrambled (scr) control. C) eggs or (E-G) L4 dCas9::VP64 transgenic C. elegans were placed on sgRNA-expressing bacteria. Lifespans were performed at 20 °C. Experiments were performed in four independent biological trials for sgRNA
col-120 and three independent biological trials for sgRNA
col-10, as shown individually here. The Mantel-Cox log-rank test was used for statistics. Data, details, and statistics of lifespan assays are in Extended Data. K) dCas9-activated expression of
col-120 enhanced the expression of
hsp-16.2 promoter-driven GFP after a heat shock at 33 °C for 2 hours with 6 hours of recovery at 20 °C, as shown in the representative fluorescent images. The bars represent mean + SEM and are a composite of three independent biological trials. The dots represent the quantified fluorescence intensity of Phsp-16.2::GFP in arbitrary units of individual C. elegans. Welch's t-test was used for significance analysis. L-O) Heat stress survival curves showed enhanced survival of animals with the dCas9-activated expression of
col-120 to high temperature (35 °C). Experiments were performed in four independent biological trials, as shown individually. The details of the heat resistance assay, summary, and statistical analysis are in Extended Data. The Mantel-Cox log-rank test was used for significance analysis.