KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| GFP Trap | ChromoTek | Cat#gtm-20 |
| Bacterial and Virus Strains | ||
| E. coli: OP50–1 | Caenorhabditis Genetics Center | RRID:WB-STRAIN:OP50–1 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Protein: Cas9-NLS purified protein | QB3 MacroLab, UC Berkley | N/A |
| Stellaris RNA FISH Hybridization buffer | LGC BioSearch Technologies | Cat#SMF-HB1–10 |
| Stellaris RNA FISH Wash Buffer A | LGC BioSearch Technologies | Cat#SMF-WA1–60 |
| Stellaris RNA FISH Wash Buffer B | LGC BioSearch Technologies | Cat#SMF-WB1–20 |
| ProLong Antifade Mountant with DAPI | Invitrogen | Cat#P36935 |
| TRIzol Reagent | Invitrogen | Cat#15596026 |
| DreamTaq DNA polymerases | Thermo Scientific | Cat#EP0705 |
| Phusion High-Fidelity DNA polymerases | Thermo Scientific | Cat#F530L |
| Protease Inhibitor Cocktail Tablet | Roche | Cat#05892970001 |
| Critical Commercial Assays | ||
| SuperScript III First-Strand Synthesis System | Invitrogen | Cat#18080051 |
| Gateway LR Clonase II Enzyme Mix | Thermo Scientific | Cat#11791–100 |
| Gibson Assembly Master Mix | New England BioLabs | Cat#E2611S |
| Deposited Data | ||
| Raw and Analyzed Data | This paper | GEO: GSE124714 |
| Experimental Models: Organisms/Strains | ||
| C. elegans: Strain wild type N2 | Caenorhabditis Genetics Center | RRID:WB-STRAIN:N2_(ancestral) |
| C. elegans: Strain Pmec-7-GFP(muIs32) II | [18] | CZ10969 |
| C. elegans: Strain Pmec-4-GFP(zdIs5) I | [18] | CZ10175 |
| C. elegans: Strain Pmec-17-GFP(uIs31) III | [62] | CZ25265 |
| C. elegans: Strain zdIs5 I; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26592 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; Pcgh-1-CGH-1::GFP(dhIs1000) | This paper | CZ26244 |
| C. elegans: Strain Pcgh-1-CGH-1::GFP(dhIs1000); Pmec-4-mKate2::CAR-1(juEx7739) | This paper | CZ25819 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(juSi338) IV; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ25293 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II | This paper | CZ11541 |
| C. elegans: Strain zdIs5 I; dcap-2(tm2470) IV | This paper | CZ25658 |
| C. elegans: Strain zdIs5 I; cgh-1(ok492) III/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III] | This paper | CZ11540 |
| C. elegans: Strain zdIs5 I; dcap-1(ok2139) IV | This paper | CZ11537 |
| C. elegans: Strain zdIs5 I; dcap-1(ok2139) dcap-2(tm2470) IV | This paper | CZ27317 |
| C. elegans: Strain zdIs5 I; patr-1(tm2402) II/mIn1 [mIs14 dpy-10(e128) II] | This paper | CZ11759 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; uIs31 III | This paper | CZ25319 |
| C. elegans: Strain muIs32 II; Pmec-4-GFP::CAR-1(juEx7280) | This paper | CZ23991 |
| C. elegans: Strain car-1(ju1505) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II | This paper | CZ25262 |
| C. elegans: Strain car-1(ju1506) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II | This paper | CZ25264 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; loxP-Pcar-1-GFP::car-1g+3’UTR-loxP (juSi343) IV | This paper | CZ25401 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ25112 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmyo-3-car-1(juEx7913) | This paper | CZ26723 |
| C. elegans: Strain Pmec-4-GFP::CAR-1 (juSi338) IV | This paper | CZ24932 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(Sm) (juSi346) IV | This paper | CZ25350 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(ΔSm) (juSi345) IV | This paper | CZ25325 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(Sm+FDF) (juSi349) IV | This paper | CZ25432 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(RGG) (juSi350) IV | This paper | CZ25433 |
| C. elegans: Strain Pmec-4-GFP::CAR-1(ΔFDF+RGG) (juSi361) IV | This paper | CZ25884 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::car-1(ΔSm) [juSi345] IV | This paper | CZ25443 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::car-1(Sm) [juSi346] IV | This paper | CZ25444 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::car-1(Sm+FDF) [juSi349] IV | This paper | CZ25566 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::car-1(RGG) [juSi350] IV | This paper | CZ25567 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-4-GFP::car-1(ΔFGF+RGG) [juSi361] IV | This paper | CZ25938 |
| C. elegans: Strain zdIs5 I; Prgef-1-GFP::car-1(juIs526) | This paper | CZ25296 |
| C. elegans: Strain zdIs5 I; Prgef-1-GFP::car-1(ΔSm) (juIs549) | This paper | CZ26634 |
| C. elegans: Strain dcap-2(tm2470) IV; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ26429 |
| C. elegans: Strain Pmec-7-tagRFP::mito(jsIs1073) | This paper | NM3573 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-7-tagRFP::mito(jsIs1073) | This paper | CZ27549 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; micu-1::GFP(ju1783) IV | This paper | CZ27519 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-7-mRFP(jsIs973) III; micu-1::GFP(ju1783) IV | This paper | CZ27538 |
| C. elegans: Strain muIs32 II; mcu-1(ju1154) IV | This paper | CZ20877 |
| C. elegans: Strain muIs32 II; micu-1(ju1155) IV/nT1 [qIs51 IV;V] | This paper | CZ26661 |
| C. elegans: Strain muIs32 II; car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; mcu-1(ju1154) IV | This paper | CZ25925 |
| C. elegans: Strain muIs32; car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; micu-1(ju1155) IV/tmC9 [In(glb-19 lgc-52 In(mec-3 unc-31)) IV] | This paper | CZ27010 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ26629 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-7-mRFP(jsIs973) III; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ26687 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; micu-1(ju1155) IV/nT1 [qIs51 IV;V]; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ26737 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; mcu-1(ju1154) IV; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ26857 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; mcu-1(ju1154) micu-1(ju1156) IV/nT1 [qIs51 IV;V]; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ26858 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-7-mRFP(jsIs973) III; ; mcu-1(ju1154) IV; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ27060 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-7-mRFP(jsIs973) III; micu-1(ju1155) IV/tmC9 [In(glb-19 lgc-52 In(mec-3 unc-31)) IV]; Pmec-4-mito::GCaMP5(juIs550) | This paper | CZ27061 |
| C. elegans: Strain Punc-17::GFP(vsIs48); [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26594 |
| C. elegans: Strain Punc-17::mCherry(nuIs321); Pcgh-1-CGH-1::GFP(dhIs1000) | This paper | CZ26802 |
| C. elegans: Strain muIs32 II; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26384 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26541 |
| C. elegans: Strain zdIs5 I; dcap-2(tm2470) IV; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26419 |
| C. elegans: Strain zdIs5 I; patr-1(tm2402) II/mIn1 [mIs14 dpy-10(e128) II]; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26425 |
| C. elegans: Strain zdIs5 I; cgh-1(ok492) III/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26511 |
| C. elegans: Strain cgh-1(ok492) III/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ25879 |
| C. elegans: Strain patr-1(tm2402) II/mIn1 [mIs14 dpy-10(e128) II]; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ26426 |
| C. elegans: Strain dcap-2(tm2470) IV; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ26429 |
| C. elegans: Strain dcap-1(ok2139) IV; Pmec-4-GFP::CAR-1(juSi338) IV | This paper | CZ26532 |
| C. elegans: Strain dcap-2(tm2470) IV; Pmec-4-GFP::CAR-1(juSi338) IV; [Pdcap-1-DCAP-1::dsRed+rol-6(su1006)](bpIs37) | This paper | CZ26632 |
| C. elegans: Strain micu-1::gfp(ju1783) IV | This paper | CZ27508 |
| C. elegans: Strain Pmec-4-GFP::DCAP-1(juEx8024) | This paper | CZ27378 |
| C. elegans: Strain zdIs5 I; mcu-1(ju1154) IV | This paper | CZ21242 |
| C. elegans: Strain zdIs5 I; micu-1(ju1155) IV/nT1 [qIs51 IV;V] | This paper | CZ27382 |
| C. elegans: Strain zdIs5 I; dcap-1(ok2139) mcu-1(ju1154) IV | This paper | CZ27368 |
| C. elegans: Strain zdIs5 I; dcap-1(ok2139)/nT1 dcap-1(ok2139) micu-1(ju1155) IV | This paper | CZ27381 |
| C. elegans: Strain muIs32 II; Pmec-7-tagRFP::mito(jsIs1073) | This paper | CZ25635 |
| C. elegans: Strain car-1(tm1753) I/hT2 [bli-4(e937) I let-?(q782) GFP(qIs48) I;III]; muIs32 II; Pmec-7-tagRFP::mito(jsIs1073) | This paper | CZ25651 |
| C. elegans: Strain muIs32 II; mcu-1(ju1154) IV; Pmec-7-tagRFP::mito(jsIs1073) | This paper | CZ26537 |
| C. elegans: Strain muIs32 II; micu-1(ju1155) IV//tmC9 [In(glb-19 lgc-52 In(mec-3 unc-31)) Pmyo-2-Venus(tmIs1221)] IV; Pmec-7-tagRFP::mito(jsIs1073) | This paper | CZ27030 |
| C. elegans: Strain zdIs5 I; Pmec-4-micu-1-gfp(juEx8048) | This paper | CZ27739 |
| C. elegans: Strain zdIs5 I; Pmec-4-micu-1-gfp(juEx8050) | This paper | CZ27756 |
| C. elegans: Strain Pmec-7-mRFP(jsIs973) III; Pmec-4-mito::GCaMP5(juIs550); Pmec-4-micu-1-gfp(juEx8048) | This paper | CZ27764 |
| C. elegans: Transgenic allele: N2 injected with 10 ng/μl of pCZGY2860 [Pmec-4-GFP::car-1(genomic)::let-858_3’UTR] and 5 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juEx7280 |
| C. elegans: Transgenic allele: N2 injected with 10 ng/μl pCZGY3372 [Pmyo-3-car-1(cDNA)::let-858_3’UTR] and 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juEx7913 |
| C. elegans: Transgenic allele: N2 injected with 10 ng/μl pCZGY3411 [Pmec-4-GFP::dcap-1(cDNA)::let-858_3’UTR] and 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juEx8024 |
| C. elegans: Transgenic allele: N2 injected with 2 ng/μl pCZGY3416 [Pmec-4-micu-1-gfp] and 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juEx8048 |
| C. elegans: Transgenic allele: N2 injected with 10 ng/μl pCZGY3416 [Pmec-4-micu-1-gfp] and 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juEx8050 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3366 [Pcar-1(1062bp upstream)-GFP::car-1(genomic+768bp 3’UTR)]. | This paper | juSi343 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3362 [Pmec-4-GFP::car-1(cDNA)::let-858_3’UTR]. | This paper | juSi338 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3367 [Pmec-4-GFP::car-1(Sm)::let-858_3’UTR]. | This paper | juSi346 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3368 [Pmec-4-GFP::car-1(ΔSm)::let-858_3’UTR]. | This paper | juSi345 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3369 [Pmec-4-GFP::car-1(Sm+FDF)::let-858_3’UTR]. | This paper | juSi349 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3370 [Pmec-4-GFP::car-1(RGG)::let-858_3’UTR]. | This paper | juSi350 |
| C. elegans: Single-copy transgenic allele: N2 injected with 50ng/μL of pCZGY3371 [Pmec-4-GFP::car-1(ΔFDF+RGG)::let-858_3’UTR]. | This paper | juSi361 |
| C. elegans: Multi-copy transgenic allele: CZ20310 [Pmex-5-his-72::miniSOG(juSi164) III unc-119(ed3) III] injected with 10 ng/μl pCZGY3363 [Prgef-1-GFP::car-1(genomic)::let-858_3’UTR] and 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juIs526 |
| C. elegans: Multi-copy transgenic allele: CZ20310 [Pmex-5-his-72::miniSOG(juSi164) III unc-119(ed3) III] injected with 10 ng/μl pCZGY3364 [Prgef-1-GFP::car-1(ΔSm)::let-858_3’UTR] 3 ng/μl pCFJ90[Pmyo-2-mCherry]. | This paper | juIs549 |
| C. elegans: Multi-copy transgenic allele: N2 injected with 40 ng/μL of Pmec-4-mito::GCaMP5 and treated with TMP/UV. | This paper | juIs550 |
| C. elegans: GFP knock-in allele: N2 injected with 50 ng/μl of pCZGY3412, 50 ng/μl of pCZGY3413, 10 ng/μl of pCZGY3414, 3 ng/μl Pmyo-2-mCherry and 3 ng/μl of Pmyo-3-mCherry. Strains were treated with heat shock as described in Dickinson et al., 2015. | This paper | ju1783 |
| Oligonucleotides | ||
| Alt-R® CRISPR-Cas9 crRNA for car-1: to make car-1(ju1505) and car-1(ju1506) deletion: /AltR1/rCrA rArGrC rUrCrG rArCrA rUrCrC rGrUrU rArCrG rGrUrU rUrUrA rGrArG rCrUrA rUrGrC rU/AltR2/ | This paper | N/A |
| Alt-R® CRISPR-Cas9 crRNA for car-1: to make car-1(ju1505) and car-1(ju1506) deletion: /AltR1/rCrU rArArU rCrUrU rGrCrU rUrCrC rGrArU rGrUrA rGrUrU rUrUrA rGrArG rCrUrA rUrGrC rU/AltR2/ | This paper | N/A |
| Alt-R® CRISPR-Cas9 crRNA for micu-1: to make micu-1(ju1155) and micu-1(ju1156) deletion: /AltR1/rGrU rGrGrG rCrGrG rUrUrU rCrUrC rGrCrG rArCrG rGrUrU rUrUrA rGrArG rCrUrA rUrGrC rU/AltR2/ | This paper | N/A |
| Alt-R® CRISPR-Cas9 crRNA for micu-1: to make micu-1(ju1155) and micu-1(ju1156) deletion: /AltR1/rArU rCrGrA rUrCrC rUrArA rCrGrA rArGrA rCrArG rGrUrU rUrUrA rGrArG rCrUrA rUrGrC rU/AltR2/ | This paper | N/A |
| Alt-R CRISPR-Cas9 tracrRNA | IDT | Cat#1072532 |
| Recombinant DNA | ||
| Plasmid: Pmec-4-GFP::car-1(genomic)-let-858_3’UTR | This paper | pCZGY2860 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1(cDNA)-let-858_3’UTR for CRISPR | This paper | pCZGY3362 |
| Plasmid: Prgef-1-GFP::car-1(cDNA)-let-858_3’UTR | This paper | pCZGY3363 |
| Plasmid: Prgef-1-GFP::car-1(ΔSm)-let-858_3’UTR | This paper | pCZGY3364 |
| Plasmid: FRT-Hygro-FRT-loxP-car-1-loxP for CRISPR | This paper | pCZGY3365 |
| Plasmid: FRT-Hygro-FRT-Pcar-1(1062bp upstream)-GFP::car-1(genomic+768bp 3’UTR) for CRISPR | This paper | pCZGY3366 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1(Sm)-let-858_3’UTR for CRISPR | This paper | pCZGY3367 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1(ΔSm)-let-858_3’UTR for CRISPR | This paper | pCZGY3368 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1-(ΔRGG)-let-858_3’UTR for CRISPR | This paper | pCZGY3369 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1(RGG)-let-858_3’UTR for CRISPR | This paper | pCZGY3370 |
| Plasmid: FRT-Hygro-FRT-Pmec-4-GFP::car-1(ΔFDF+RGG)-let-858_3’UTR for CRISPR | This paper | pCZGY3371 |
| Plasmid: Pmyo-3-car-1(cDNA)-unc-54_3’UTR | This paper | pCZGY3372 |
| Plasmid: Pmec-4-GFP::dcap-1(cDNA)-let-858_3’UTR | This paper | pCZGY3411 |
| Plasmid: Peft-3::Cas9+micu-1_C1_sgRNA | This paper | pCZGY3412 |
| Plasmid: Peft-3::Cas9+micu-1_C2_sgRNA | This paper | pCZGY3413 |
| Plasmid: GFP-SEC-3xFLAG_micu-1_C1_homology_arm | This paper | pCZGY3414 |
| Plasmid: Pmec-4-micu-1-gfp | This paper | pCZGY3416 |
| Software and Algorithms | ||
| ImageJ | NIH image | RRID:SCR_003070 |
| GraphPad Prism 7 | GraphPad Software, Inc. | RRID:SCR_002798 |
| ZEN | Zeiss | https://www.zeiss.com/microscopy/us/downloads/zen.html |