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Experimental Models
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| Bacterial strains |
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| Escherichia coli clone for C. elegans SMD-1 F47G4.7 |
RNAi feeding library (Open Biosystems, Huntsville, AL) |
N/A |
| Escherichia coli clone for C. elegans KLP-7 K11D9.1 |
RNAi feeding library (Open Biosystems, Huntsville, AL) |
N/A |
| Escherichia coli clone for C. elegans ZYG-9 F22B5.7 |
RNAi feeding library (Ahringer RNAi library) |
N/A |
| Escherichia coli clone for C. elegans TBG-1 F58A4.8 |
RNAi feeding library (Open Biosystems, Huntsville, AL) |
N/A |
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C. elegans strains
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C. elegans wild type isolate |
CGC |
N2 |
| unc-119(ed3) III; ltIs76 [pAA178: pie-1p::mCHERRY::sp12 + unc-119 (+)]; ltIs25 [pAZ132; pie-1p::GFP::tba-2 + unc-119 (+)] |
Audhya lab |
MSN146 |
|
ebp-2(or1954[ebp-2::mKate2]) II; mTurquoise2::H2B (his-72::linker::mTurquoise2) III (CRISPR); unc-119(ed3); ojIs23 [SP12::GFP + unc-119(+)]
|
(Maheshwari et al., 2023) |
OCF163 |
|
spd-5(vie26[gfp::spd-5 +loxP]) I; unc-119(ed3); ocfIs2[pie-1 prom::mCherry::SP12::pie-1 3’UTR +unc119 (+)];mTurquoise2::H2B (his-72::linker::mTurquoise2) III
|
(Maheshwari et al., 2023) |
OCF176 |
|
unc-119(ed3); ocfIs2[pie-1 prom::mCherry::SPl2::pie-1 3’UTR +unc119 (+)];ltIs25 [pAZ132; pie-1p::GFP::tba-2 + unc-119 (+)]; mTurquoise2::H2B (his-72::linker::mTurquoise2) III
|
(Maheshwari et al., 2023) |
OCF181 |
|
unc-119(ed3); ocfIs2[pie-1 prom::mCherry::SP12::pie-1 3’UTR +unc119 (+)];ltIs25 [pAZ132; pie-1p::GFP::tba-2 + unc-119 (+)]; mTurquoise2::H2B (his-72::linker::mTurquoise2) III; ocf101[atln-1::3xFLAG::degron] IV (CRISPR); TIR1::mRuby IV
|
(Maheshwari et al., 2023) |
OCF183 |
|
unc-119(ed3); ojIs23 [SP12::GFP + unc-119(+)]; itSi569Itbg-1::mCherry); mTurquoise2::H2B (his-72::linker::mTurquoise2) III
|
(Maheshwari et al., 2023) |
OCF184 |
|
unc-119(ed3); ocfIs2[pie-1 prom::mCherry::SP12::pie-1 3’UTR +unc119 (+)]; cb-unc-119(+)]I; ltSi1141[pOD1021/pVV103; Pspd-2::GFP::SPD-5 reencoded; cb-unc-119(+)]II; unc-119(ed3) III
|
This study |
OCF187 |
|
unc-119(ed3); ocfIs2[pie-1 prom::mCherry::SP12::pie-1 3’UTR +unc119 (+)]; ltSi592[pVV168; Pspd-2::GFP-spd-5 mut 653,658, reencoded; cb-unc-119(+)]II; unc-119(ed3)III
|
This study |
OCF189 |
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unc-119(ed3); ojIs23 [SP12::GFP + unc-119(+)]; PH::gfp; tba-1(pg77[tba-1::TagRFP-T + loxP]) I; uIs31 III
|
This study |
OCF193 |
|
ltSi592[pVV168; Pspd-2::GFP::spd-5 mut 653,658, reencoded; cb-unc-119(+)]II; unc-119(ed3)III; spd-5(wow36[tagrfp-t^3xmyc::spd-5]) I
|
This study |
OCF200 |
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Pspd-2::GFP::SPD-5 reencoded; cb-unc-119(+)]II; unc-119(ed3) III; spd-5(wow36[tagrfp-t^3xmyc::spd-5]) I
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This study |
OCF201 |
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tba-1(pg77[tba-1::TagRFP-T + loxP]) I; uIs31 III; Pspd-2::GFP::SPD-5 reencoded; cb-unc-119(+)]II; unc-119(ed3) III
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This study |
OCF212 |
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tba-1(pg77[tba-1::TagRFP-T + loxP]) I; uIs31 III; ltSi592[pVV168; Pspd-2::GFP::spd-5 mut 653,658, reencoded; cb-unc-119(+)]II; unc-119(ed3)III
|
This study |
OCF213 |
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unc-119(ed3); ojIs23 [SP12::GFP + unc-119(+)];tba-1(pg77[tba-1::TagRFP-T + loxP]) I; ltSi1219[pMO104; Pspd-2::SPD-5 (re-encoded) (WT, untagged);cb-unc-119(+)]II; unc-119(ed3)III
|
This study |
OCF214 |
|
unc-119(ed3); ojIs23 [SP12::GFP + unc-119(+)]; tba-1(pg77[tba-1::TagRFP-T + loxP]) I; ItSi1219[pMO104; Pspd-2::spd-5 S653A S658A::spd-5 3′UTR; cb-unc119(+)]II; unc-119(ed3)III
|
This study |
OCF215 |
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Recombinant DNA
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| Vector for expressing dsRNA: pL4440 |
Addgene |
# 1654 |
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Antibodies
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| None |
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Oligonucleotides and other sequence-based reagents
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| TAATACGACT CACTATAGGT GGAATT GTCCGCTACTGAT |
IDT |
spd-5 RNAi F |
| AATTAACCCT CACTAAAGGT GTATT C AACGAGTGCCTGA |
IDT |
spd-5 RNAi R |
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Chemicals, Enzymes and other reagents
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| Amplicillin |
Sigma Aldrich |
Cat # A9518 |
| Indole-3-acetic acid (auxin) |
Alfa Aesar |
Cat # A10556 |
| IPTG |
Sigma Aldrich |
Cat # I6758 |
| Qiagen MinElute Reaction Cleanup Kit Qiagen Cat |
Qiagen |
Cat # 28206 |
| MEGAscript™ T7 Transcription Kit |
Invitrogen |
Cat # AM1333 |
| MEGAscript™ T3 Transcription Kit |
Invitrogen |
Cat # AM1338 |
| Phenol:CHCl3:Isoamyl Alcohol |
Invitrogen |
Cat # 15593031 |
| Ethanol 200 proof |
The Warner-Graham company |
Cat # 64-17-5 |
| Agarose |
Invitrogen |
Cat # 214010 |
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Software
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| FIJI (ImageJ release 2.1.0) |
(Schindelin et al, 2012) |
https://imagej.nih.gov/ij/
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| GraphPad Prism [Version 10.1.1 (270)] |
GraphPad |
https://www.graphpad.com/scientificsoftware/prism/
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| Nikon Elements Software |
Nikon USA |
N/A |
| IMOD or 3dmod [version 4.11.1] |
https://bio3d.colorado.edu
|
N/A |
| AMIRA 6.5.0 (release 2018-03-07) |
Thermo Fisher Scientific |
N/A |
| Amira ZIB edition Version 2020.1 |
Zuse Institute Berlin Dep. Visual Data Analysis, Takustr.7, Berlin, Germany. |
N/A |
| Adobe Photoshop CC (release 23.1) |
Adobe |
N/A |
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Other
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