Figure 5.

A functional UPR^mt pathway in neurons is required for peripheral induction of UPR^mt. (A) Targeting sequences of UPR^mt pathway genes for CRISPR-Cas9 knockout, together with their PAM sequences. (B) Deletions of UPR^mt pathway genes produced by CRISPR/Cas9 are detected by T7E1 assay. Representative DNA gels of T7E1 assay reveals the PCR products amplified from genomic DNA of control worms, or worms with UPR^mt pathway gene deletion in the nervous system. (C, D) Neural knockout of atfs-1 or dve-1 suppresses the UPR^mt in distal tissues in rab-3p::Cas9+u6p::spg-7-sg worms. Representative fluorescent images (C) and quantification of hsp-6p::GFP reporter expression (D) in animals containing rab-3p::Cas9+u6p::spg-7-sg (control) or rab-3p::Cas9+u6p::spg-7-sg with neural-specific knockout of UPR^mt genes are shown. mec-7p::RFP is used as co-injection marker. n ≥ 28, error bars indicate mean ±SE. A Student's t-test is used to assess significance: ^*P < 0.05. (E) Neural expression of ATFS-1^Δ1-32 induces UPR^mt in distal tissues when worms reach day 1 adult stage. odr-1 p::dsRed is used as co-injection marker.