Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Drosophila melanogaster) |
prp4 | NA | FLYB:FBgn0027587 | |
| Gene (Drosophila melanogaster) |
timeless (tim) | NA | FLYB:FBgn0014396 | |
| Gene (Drosophila melanogaster) |
period (per) | NA | FLYB:FBgn0003068 | |
| Gene (Drosophila melanogaster) |
prp8 | NA | FLYB:FBgn0033688 | |
| Gene (Drosophila melanogaster) |
brr2 | NA | FLYB:FBgn0263599 | also known as l(3)72Ab |
| Gene (Drosophila melanogaster) |
prp3 | NA | FLYB:FBgn0036915 | |
| Gene (Drosophila melanogaster) |
prp31 | NA | FLYB:FBgn0036487 | |
| Strain, strain background (Drosophila melanogaster) |
iso31 | from laboratory stocks |
NA | |
| Genetic reagent (Drosophila melanogaster) |
per01 | Bloomington Drosophila Stock Center (BDSC) |
FLYB:FBal0013649 | |
| Genetic reagent (Drosophila melanogaster) |
tim0 | BDSC | FLYB:FBal0035778 | |
| Genetic reagent (Drosophila melanogaster) |
TUG
(Tim-UAS-Gal4) |
BDSC | FLYB:FBtp0011839 | |
| Genetic reagent (Drosophila melanogaster) |
pdfGal4;
pdfG4 |
BDSC | FLYB:FBtp0011844 | |
| Genetic reagent (Drosophila melanogaster) |
elavGal4;
elavG4 |
BDSC | BDSC:25750 | |
| Genetic reagent (Drosophila melanogaster) |
GMRGal4;
GMR |
BDSC | FLYB:FBti0002994 | |
| Genetic reagent (Drosophila melanogaster) |
prp4RNAi(GD) | Vienna Drosophila Resource Center (VDRC) |
VDRC:27808 | |
| Genetic reagent (Drosophila melanogaster) |
prp4RNAi(KK) | VDRC | VDRC:107042 | |
| Genetic reagent (Drosophila melanogaster) |
prp8RNAi(GD) | VDRC | VDRC:18565 | |
| Genetic reagent (Drosophila melanogaster) |
prp3RNAi(GD) | VDRC | VDRC:25547 | |
| Genetic reagent (Drosophila melanogaster) |
prp3RNAi(KK) | VDRC | VDRC:103628 | |
| Genetic reagent (Drosophila melanogaster) |
prp31RNAi(KK) | VDRC | VDRC:103721 | |
| Genetic reagent (Drosophila melanogaster) |
brr2RNAi(KK) |
VDRC | VDRC:110666 | |
| Genetic reagent (Drosophila melanogaster) |
prp82e1 | BDSC | FLYB: FBal0190235; BDSC:25905 |
|
| Genetic reagent (Drosophila melanogaster) |
prp82e2 | BDSC | FLYB:FBal0190015; BDSC:25912 |
|
| Genetic reagent (Drosophila melanogaster) |
brr2e03171 | BDSC | FLYB:FBti0041681; BDSC:18127 |
|
| Genetic reagent (Drosophila melanogaster) |
UAS-Dicer2;
Dcr2 |
BDSC | FLYB:FBtp0036672 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-tim-spliced; tim-spliced |
this paper | NA | generated by the site-specific PhiC31 Integration System (Rainbow Transgenics) using the attP on the 3rd chromosome; pUAST-tim-spliced plasmid was used for injection |
| Genetic reagent (Drosophila melanogaster) |
UAS-tim-retained; tim-retained |
this paper | NA | generated by the site-specific PhiC31 Integration System (Rainbow Transgenics) using the attP on the 3rd chromosome; pUAST-tim- retained plasmid was used for injection |
| Genetic reagent (Drosophila melanogaster) |
UAS-tim-retained+ssM; tim-retained+ ssM |
this paper | NA | generated by the site-specific PhiC31 Integration System (Rainbow Transgenics) using the attP on the 3rd chromosome; pUAST-tim- retained+ssM plasmid was used for injection |
| Cell line (Drosophila melanogaster) |
S2 | ATCC (Manassas, VA) |
FLYB:FBtc0000181; RRID:CVCL:Z992 |
|
| Antibody | guinea pig anti- PER (UP1140) |
Garbe et al., 2013 | NA | 1:1000 |
| Antibody | rat anti- TIM (UPR42) |
Jang et al., 2015 | NA | 1:1000 |
| Antibody | rabbit anti-PDF (HH74) |
Garbe et al., 2013 | NA | 1:500 |
| Antibody | mouse anti-LaminC |
Developmental Studies Hybridoma Bank (DSHB) |
LC28.26 | 1:500 |
| Antibody | mouse anti-HSP70 |
Sigma | Cat# H5147 | 1:5000 |
| Recombinant DNA reagent |
pIZ/V5-His plasmid |
ThermoFischer | Cat# V800001 | backbone |
| Recombinant DNA reagent |
pBluescript-tim | lab collection | NA |
tim sequence contained tim-tiny; used for subcloning |
| Recombinant DNA reagent |
tim-spliced; pIZ-tim-spliced |
this paper | NA |
tim cDNA was subcloned into pIZ-V5 plasmids |
| Recombinant DNA reagent |
tim-retained; pIZ-tim-retained |
this paper | NA |
tim-tiny intron was subcloned into pIZ-tim- spliced vector from pBluescript-tim plasmid |
| Recombinant DNA reagent |
tim-retained+ ssM; pIZ-tim-retained+ssM |
this paper | NA | generated by mutagenesis of the 5’ splice donor site of tim-tiny intron from pIZ-tim-retained plasmid |
| Recombinant DNA reagent |
tim-spliced; pUAST-tim-spliced |
this paper | NA |
tim cDNA was subcloned from pIZ-tim- spliced into pUAST -attB vector |
| Recombinant DNA reagent |
tim-retained; pUAST-tim-retained |
this paper | NA |
tim cDNA was subcloned from pIZ-tim-retained into pUAST-attB vector |
| Recombinant DNA reagent |
tim-retained + ssM; pUAST-tim-retained+ssM |
this paper | NA |
tim cDNA was subcloned from pIZ-tim- retained+ssM into pUAST-attB vector |
| Sequence- based reagent |
tim PP11542 (‘mRNA’) _F |
ATGGACTGGTTACTAGCAACTCC | ||
| Sequence- based reagent |
tim PP11542 (‘mRNA’) _R |
GGTCCTCATAGGTGAGCTTGT | ||
| Sequence- based reagent |
per_F | CGTCAATCC ATGGTCCCG |
||
| Sequence- based reagent |
per_R | CCTGAAAGACGCGATGGTG | ||
| Sequence- based reagent |
clk_F | GGATGCCAATGCCTACGAGT | ||
| Sequence- based reagent |
clk_R | ACCTACGAAAGTAGCCCACG | ||
| Sequence- based reagent |
prp4_F | CACAAGCAGCATCTTTGTATGG | ||
| Sequence- based reagent |
prp4_R | TGTGGAGTCCCACATTCTTG | ||
| Sequence- based reagent |
tim-tiny_retained_F | AAACGTGAGTTAAAGTCAACC | ||
| Sequence- based reagent |
tim-tiny_retained_R | GAGAGGCACACAGCATATC | ||
| Sequence- based reagent |
tim-tiny_spliced_F | CCGCTGGACAAACTCAACCTC | ||
| Sequence- based reagent |
tim-tiny_spliced_R | TCGGTATCGCCGAGATCCACG | ||
| Sequence- based reagent |
tim-cold_retained_F | GGCTCATGATCATTGCAGCAGC | ||
| Sequence- based reagent |
tim-cold_retained_R | ATAGTGGGGCACCCGGATCTC | ||
| Sequence- based reagent |
tim-cold_spliced_F | TTAAACAGCGACAATGTCTCTTTGG | ||
| Sequence- based reagent |
tim-cold_spliced_R | GAATTGGATCCTCAGTGATAGTGGG | ||
| Sequence- based reagent |
tim_non_spanning ('exon')_F |
GAAGAACAACGATATTGTGGGAAAG | ||
| Sequence- based reagent |
tim_non_spanning ('exon')_R |
AGTGGGAGTTGTCAGCAAAG | ||
| Sequence- based reagent |
per_retained_F | GAGGACCAGACACAGCACGG | ||
| Sequence- based reagent |
per_retained_R | CGGAGGCAATTGCTCACTCGT | ||
| Sequence- based reagent |
per_spliced_F | GAGGACCAGACACAGCACGG | ||
| Sequence- based reagent |
per_spliced_R | TCGCGTTGATTCGAAGAATCGTT | ||
| Sequence- based reagent |
rp49_F | GACGCTTCAAGGGACAGTATCTG | ||
| Sequence- based reagent |
rp49_R | AAACGCGGTTCTGCATGAG | ||
| Sequence- based reagent |
tim_tinySSdonorT > A_F | CTGGACAAACGAGAGTTAAAGTCAACC | ||
| Sequence- based reagent |
tim_tinySSdonorT > A_R | CGGTCCCAGCTTTTTGGC | ||
| Commercial assay or kit |
RNeasy Plus Mini Kit |
Qiagen | Cat# 74134 | |
| Commercial assay or kit |
Superscript II Reverse Transcriptase |
ThermoFischer | Cat# 18064014 | |
| Commercial assay or kit |
TRIzol Reagent | ThermoFischer | Cat# 15596026 | |
| Commercial assay or kit |
Q5 Site-Directed Mutagenesis Kit |
NEB | Cat# E0554S | |
| Commercial assay or kit |
Effectene Transfection Reagent |
Qiagen | Cat# 301425 | |
| Software, algorithm |
Graphpad Prism v7 |
Graphpad Software |
https://www.graphpad.com/ | |
| Software, algorithm |
JTK_CYCLE v3 | Hughes et al., 2010 | NA | |
| Software, algorithm |
ImageJ | NIH | https://imagej.nih.gov/ij/ | |
| Software, algorithm |
ClockLab Software |
Actimetrics (Wilmette, IL) |
https://actimetrics.com/products/clocklab |