KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-CRHBP Antibody | Santa Cruz | sc-20630 |
| Mouse monoclonal anti-beta-actin Antibody | ABGENT | AM1829B |
| Mouse monoclonal anti-EGFP antibody | Laboratory of Nathaniel Heintz | N/A |
| Chicken anti-EGFP antibody | AVES | GFP-1020 |
| Sheep anti-DIG polyclonal antibodies | Roche | 11333089001 |
| Biotinylated Rabbit anti-sheep IgG antibody | Vector lab | BA-6000 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Oxytocin acetate salt | BACHEM | H-2510.0025 |
| CRF (human, rat) acetate salt | BACHEM | H-2435.0001 |
| CRF (6-33) (human, rat) | BACHEM | H-3456.1000 |
| Picrotoxin | TOCRIS | 1128 |
| D-AP5 | TOCRIS | 0106 |
| CNQX disodium salt | TOCRIS | 1045 |
| Polyethylenimine HCI MAX | Polysciences, Inc. | 24765-2 |
| Critical Commercial Assays | ||
| TSA plus cyanine 3 system | PerkinElmer | NEL744001KT |
| RNAeasy Mini Kit | QIAGEN | 74106 |
| LightCycler 480 SYBR Green I Master | Roche | 04887352001 |
| DIG RNA Labeling Kit (SP6/T7) | Roche | 11175025910 |
| Dynabeads MyOne Streptavidin T1 | Invitrogen | 65601 |
| RNeasy Plus Micro Kit | QIAGEN | 74034 |
| Ovation RNA-seq System V2 | Nugen | 7102 |
| TruSeq Nano DNA LT kit | illumina | 15041757 |
| C&B Metabond | parkell | S380 |
| SuperScript IV First-Strand Synthesis System | Invitrogen | 18091050 |
| NuPAGE 4-12% Bis-Tris Protein Gels | Invitrogen | NP0321BOX |
| Deposited Data | ||
| No | ||
| Experimental Models: Cell Lines | ||
| 293TN producer cell line | System Biosciences | LV900A-1 |
| Experimental Models: Organisms/Strains | ||
| C57BL/6J | The Jackson Laboratory | 000664 |
| Oxtr-cre ON82 | Laboratory of Nathaniel Heintz (Nakajima N et al.,2014) | N/A |
| Rosa26-flox-EGFP-L10a | The Jackson Laboratory | 022367 |
| Oxtr-EGFP-L10a | Laboratory of Nathaniel Heintz | N/A |
| Oxtrflox/flox | The Jackson Laboratory | 008471 |
| Recombinant DNA | ||
| AAV5-EF1a-DIO-hChR2(H134R)-mCherry | University of Pennsylvania vector core | AAV-5-20297P |
| AAV8-hSyn-GFP-Cre | University of North Carolina vector Core | N/A |
| AAV8-CMV-GFP | University of North Carolina vector Core | N/A |
| Sequence-Based Reagents | ||
| Crhbp probe for ISH Forward: CCCTCTCCTGCTCTTCAGTG Reverse: ATGGCTCCAGCTGACGATAC |
This paper | N/A |
| pCMV-Crhbp cDNA (Myc-DDK-Tagged) | Origene | MR204660 |
| pSico | Ventura A et al.,2004 ( Addgene ) | 11578 |
| 19-nt sequence used for knockdown of Crhbp mRNA GAGCCATTCGAGCTAGAAA | This paper | Generated by pSicoOligomaker |
| Sense oligo containing Crhbp shRNA: TGAGCCATTCGAGCTAGAAATTCAAGAGATTTCTAG CTCGAATGGCTCTTTTTTC Antisense oligo containing Crhbp shRNA: TCGAGAAAAAAGAGCCATTCGAGCTAGAAATCTCTT GAATTTCTAGCTCGAATGGCTCA |
This paper | Generated by pSicoOligomaker |
| RT-PCR primers of Oxtr Forward: CTTCCTCAGATTCCACACCTG Reverse: TTCCAGAACATTCAGCTCCAG |
This paper | N/A |
| RT-PCR primers of CRH Forward: AAAGAAGAGAAAGGAGAAGAGGAAG Reverse: CCGCAGCCGCATGTTAG |
This paper | N/A |
| RT-PCR primers of CrhR1 Forward: CAGCCGCCTACAACTACTTCC Reverse: GGTGGAGTACGTGAGTACGATG |
This paper | N/A |
| RT-PCR primers of Crbhp Forward: TGATGCCCTTAGCAGACCTGT Reverse: CACACGGTTAATGTGTTTCCCA |
This paper | N/A |
| RT-PCR primers of GAPDH Forward: CATGGCCTTCCGTGTTCCTA Reverse: GCCTGCTTCACCACCTTCTT |
This paper | N/A |
| Software and Algorithms | ||
| MainScript | Laboratory of Nathaniel Heintz | https://gensat.rockefeller.edu/heintzp30/Bioinformatics_Flow_Chart.jsp |
| Other | ||
| Double guide canula | Plastic one | C235GS-5-0.5/SP |
| Fiber-optic cannula | Doric lenses | MFP_200/230-0.37 _1m_ZF1.25_FLT |