Trc library

Manufactured by Merck Group

Sourced in United States

The TRC library is a collection of small molecules used for high-throughput screening in pharmaceutical research and development. The core function of the TRC library is to provide a diverse set of chemical compounds that can be tested against target proteins or pathways to identify potential drug candidates.

Automatically generated - may contain errors

Lab products found in correlation

Plko 1 vector from the, by Merck Group (2 mentions) In fusion, by Takara Bio (2 mentions) Fasudil, by Selleck Chemicals (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) 12 well plate, by Corning (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Afatinib, by Selleck Chemicals (1 mentions) Polybrene, by Merck Group (1 mentions)

Spelling variants of this product

Mission TRC shRNA library (23 citations) MISSION TRC library (15 citations) MISSION TRC-Hs1.0 library (4 citations) TRC shRNA library (3 citations) TRC genome-wide shRNA library (3 citations) MISSION TRC-Mm 1.0 shRNA library (2 citations) TRC-Hs1.0 library (2 citations) MISSION® TRC version 1 shRNA library (2 citations) MISSION TRC shRNA human library (2 citations)

9 protocols using trc library

Evaluating EGFR and ROCK Inhibitors in Breast Cancer Cell Lines

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hek279T, MDA-MB-231, LM2, BT549, Cal51, Cal120 cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 9% fetal bovine serum (Sigma), 2mM glutamine, 0.1mg/ml penicillin and 0.1ml/ml streptomycin (Gibco). HCC1806, HCC38, Hs578T cells were maintained in RPMI supplemented with glutamine.
Hek293T cells were used for lentivirus production as described previously [19 (link)]. shRNAs targeting EGFR and ROCK were obtained from the TRCs1.0 library and were as follows: shEGFR-1: TRCN0000121068, shEGFR-2: TRCN0000010329, shEGFR-3: TRCN0000121206, shEGFR-4: TRCN0000121203, shROCK1-1: TRCN0000002163, shROCK1-2: TRCN0000121316, shROCK1-3: TRCN0000121095, shROCK1-4: TRCN0000002161 (TRC Library, Sigma).
For long-term cell growth assays, cells were seeded on 6-well or 12-well plates (Corning). Drugs were added on the following day and media was refreshed every third day with new compound dilutions. At the end time point, the cells were stained with crystal violet. ROCK inhibitors used were GSK269962A (Axon) and Fasudil (Selleck). EGFR inhibitors used were Gefitinib (MedChem) and Afatinib (Selleck).
For DNA content and cell cycle analysis, sub-confluent cells were incubated with 10uM Bromdeoxyuridine (BrdU) for 1.5 hours, trypsinized, fixed in 70% ice-cold ethanol, and stained with anti-BrdU and Propidium Iodide (PI).

Iskit S., Lieftink C., Halonen P., Shahrabi A., Possik P.A., Beijersbergen R.L, & Peeper D.S. (2016). Integrated in vivo genetic and pharmacologic screening identifies co-inhibition of EGRF and ROCK as a potential treatment regimen for triple-negative breast cancer. Oncotarget, 7(28), 42859-42872.

+ Open protocol

+ Expand

Lentiviral shRNA and Bidirectional cDNA Constructs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lentiviral shRNA constructs used the pLKO.1 vector from the TRC library (Sigma-Aldrich) or were produced by inserting shRNA sequences as described (Addgene http://www.addgene.org/tools/protocols/plko/). TRC library designations and target sequences are: shMDM2-1:TRCN0000003380, 5′-CTCAGCCATCAACTTCTAGTA; shMDM2-2:TRCN0000003377, 5′-GATTCCAGAGAGTCATGTGTT; shMDM4-1:TRCN0000003858, 5′-GTTCACTGTTAAAGAGGTCAT; shMDM4-2:TRCN0000003857, 5′-CACCTAGAAGTAATGGCTCAA, shMYCN-1:TRCN0000020695, 5′-CAGCAGCAGTTGCTAAAGAAA; shMYCN-2:TRCN0000020696, 5′-CGGACGAAGATGACTTCTACT; shp53:TRCN0000003753, 5′-CGGCGCACAGAGGAAGAGAAT, shp27:TRCN0000039930, 5′-GCGCAAGTGGAATTTCGATTT. The Bidirectional Neomycin resistance (BN) lentiviral cDNA expression vector utilizes the EF1α enhancer-promoter to drive a gene of interest and a CMV minimal promoter to drive Neo^r as described.^{2 (link)} BN-MYCN was produced by inserting MYCN cDNA sequences 729–2152 (NM_001293228.1, isoform 1) into the BN PshAI site using In-Fusion (Clontech).

Qi D.L, & Cobrinik D. (2016). MDM2 but not MDM4 promotes retinoblastoma cell proliferation through p53-independent regulation of MYCN translation. Oncogene, 36(13), 1760-1769.

+ Open protocol

+ Expand

Lentiviral Knockdown of Epigenetic Regulators

Check if the same lab product or an alternative is used in the 5 most similar protocols

To produce the lentiviruses of interest, HEK293T cells were transiently co-transfected with pLKO.1-shRNA encoding shRNAs targeting Suv4-20h1, Suv4-20h2 or Smad4 (TRC library, Sigma), as well as assistant vectors psPAX and pMD2.G. The culture medium was replaced with DMEM supplemented with 30% FBS 18 h later, and viral supernatants were collected 24 h later. Cells were infected with the packaged viruses with 5 μg/mL polybrene (M&C Gene Technology, MC032, China). 36–48 h after infection, cells were selected in 3 μg/mL of puromycin (M&C Gene Technology, MA009, China) for 24–36 h, after which they were plated into different dishes. Cells were then continuously selected in 1 μg/mL of puromycin for 4–5 days until no dead cells were found in the dishes.

Lyu G., Guan Y., Zhang C., Zong L., Sun L., Huang X., Huang L., Zhang L., Tian X.L., Zhou Z, & Tao W. (2018). TGF-β signaling alters H4K20me3 status via miR-29 and contributes to cellular senescence and cardiac aging. Nature Communications, 9, 2560.

+ Open protocol

+ Expand

Lentiviral and Retroviral Knockdown of STAT3 and IL-8

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lentivirus production and subsequent transduction of cells was carried out as described earlier67 (link). The following shRNA constructs (Sigma Aldrich mission TRC library) were used: shRNA against STAT3 (TRCN0000071456) and control scrambled shRNA (SHC002). The functionality of the shRNAs was validated by immunoblot. Transduced cells were selected for puromycin resistance before further analysis. Within a different approach we used two STAT3-specific shRNAs (#336, #946) and a control shRNA (#713) and cloned them into a retroviral tetracycline-inducible expression vector68 (link) containing the optimized miR-E backbone69 (link). Retroviral production and subsequent transduction of cells were performed as described earlier68 (link). Transduced cells were selected for neomycin resistance and doxycycline was used to induce shRNA expression. Cells were sorted for green fluorescent protein and dsRed prior usage. Short hairpin-mediated knockdown of IL-8 was performed by cloning IL-8-specific shRNA (#1255) into a lentiviral expression vector containing the optimized miR-E backbone69 (link). Lentiviral production, transduction and selection with neomycin were performed as described69 (link). Cells were sorted for green fluorescent protein in addition to antibiotic selection prior usage.

Grabner B., Schramek D., Mueller K.M., Moll H.P., Svinka J., Hoffmann T., Bauer E., Blaas L., Hruschka N., Zboray K., Stiedl P., Nivarthi H., Bogner E., Gruber W., Mohr T., Zwick R.H., Kenner L., Poli V., Aberger F., Stoiber D., Egger G., Esterbauer H., Zuber J., Moriggl R., Eferl R., Győrffy B., Penninger J.M., Popper H, & Casanova E. (2015). Disruption of STAT3 signalling promotes KRAS-induced lung tumorigenesis. Nature Communications, 6, 6285.

+ Open protocol

+ Expand

Lentiviral Knockdown of CBS and CTH

Check if the same lab product or an alternative is used in the 5 most similar protocols

For generating CBS and CTH knockdown in U1 and J1.1 cells, we used validated pooled gene-specific shRNAs from the RNAi Consortium (TRC) library (Sigma-Aldrich, USA; shRNA sequences given in Supplementary file 1a). The lentiviral particles were generated in HEK293T cells using the packaging vectors, psPAX2 and pMD2.G. The pLKO.1-puro vector encoding a non-mammalian targeting shRNA (shNT) was used as a control. The U1 cells were transduced with lentiviral particles in opti-MEM containing polybrene (10 μg/ml) for 6 hr. Cells were washed and stable clones were selected in culture medium containing 250 ng/ml of puromycin. Total RNA or cell lysates were prepared to validate knockdown of CBS and CTH.

Pal V.K., Agrawal R., Rakshit S., Shekar P., Murthy D.T., Vyakarnam A, & Singh A. (2021). Hydrogen sulfide blocks HIV rebound by maintaining mitochondrial bioenergetics and redox homeostasis. eLife, 10, e68487.

+ Open protocol

+ Expand

Lentiviral shRNA and Bidirectional cDNA Constructs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Qi D.L, & Cobrinik D. (2016). MDM2 but not MDM4 promotes retinoblastoma cell proliferation through p53-independent regulation of MYCN translation. Oncogene, 36(13), 1760-1769.

+ Open protocol

+ Expand

Lentiviral shRNA-mediated STAT3 knockdown

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lentivirus production and subsequent transduction of cells were carried out as described previously [25] (link). The following shRNA constructs (Sigma Aldrich Mission TRC Library) were used: shRNA against STAT3 (TRCN0000071456) and control scrambled shRNA (SHC002). The functionality of the shRNAs was validated by quantitative reverse transcription–polymerase chain reaction (PCR) analysis. Transduced cells were selected for puromycin resistance before further analysis.

Schütz A., Röser K., Klitzsch J., Lieder F., Aberger F., Gruber W., Mueller K.M., Pupyshev A., Moriggl R, & Friedrich K. (2015). Lung Adenocarcinomas and Lung Cancer Cell Lines Show Association of MMP-1 Expression With STAT3 Activation. Translational Oncology, 8(2), 97-105.

+ Open protocol

+ Expand

Lentiviral Knockdown of GPI-80 and ITGAM

Check if the same lab product or an alternative is used in the 5 most similar protocols

pLKO lentiviral vectors from the TRC library (Sigma) were produced and transduced in fetal liver CD34⁺ cells as described in supplemental material. Two different shRNAs with confirmed knockdown of GPI-80 or ITGAM were used: TRCN0000158666 (GPI-80 1), TRCN0000161721 (GPI-80 2), TRCN0000029051 (ITGAM 1), and TRCN0000029050 (ITGAM 2).

Prashad S., Calvanese V., Yao C., Kaiser J., Wang Y., Sasidharan R., Crooks G., Magnusson M, & Mikkola H. (2014). GPI-80 Defines Self-renewal Ability in Hematopoietic Stem Cells During Human Development. Cell stem cell, 16(1), 80-87.

+ Open protocol

+ Expand

Lentiviral Transduction of MPNST Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MPNST cells were transduced with lentiviral particles at 50–60% confluence. Short hairpins (sh) RNAs targeting gene of interest and control non-targeting construct were from the Sigma Aldrich TRC library. The CCHMC Viral Vector Core produced virus using a 4-plasmid packaging system (http://www.cincinnatichildrens.org/research/div/exphematology/translational/vpf/vvc/default. html). Lentiviral particles were incubated with MPNST cells in the presence of polybrene (8 μg/mL; Sigma) for 24 h, followed by selection in 2 μg/mL puromycin, which killed uninfected cells within 3 days.

Patel A.V., Chaney K.E., Choi K., Largaespada D.A., Kumar A.R, & Ratner N. (2016). An ShRNA Screen Identifies MEIS1 as a Driver of Malignant Peripheral Nerve Sheath Tumors. EBioMedicine, 9, 110-119.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!