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Drosha

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DROSHA is a protein complex that plays a critical role in the initial processing of microRNA (miRNA) precursors. It is a ribonuclease III enzyme that cleaves the primary miRNA transcript (pri-miRNA) to produce the precursor miRNA (pre-miRNA).

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Ripa buffer, by Thermo Fisher Scientific (4 mentions) Gapdh, by Santa Cruz Biotechnology (3 mentions) Taqman advanced mirna cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Powerup sybr green master mix, by Thermo Fisher Scientific (2 mentions) Polyvinylidene difluoride pvdf membrane, by Cytiva (1 mentions) Antibodies raised against, by Cell Signaling Technology (1 mentions) Hemin, by Merck Group (1 mentions) Phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) β actin, by Thermo Fisher Scientific (1 mentions) Dicer, by Santa Cruz Biotechnology (1 mentions) Sparc, by Santa Cruz Biotechnology (1 mentions) High pure mirna isolation kit, by Merck Group (1 mentions) High capacity reverse transcriptions kit, by Thermo Fisher Scientific (1 mentions) Gapmer, by Qiagen (1 mentions) Tween 20, by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Hdac1, by Abcam (1 mentions) Ddx17, by Abcam (1 mentions) Ha 11, by BioLegend (1 mentions)

4 protocols using drosha

1

Protease Inhibitor Cocktail Protocol

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Protease inhibitor cocktail, Tween-20, Hemin, High Pure miRNA Isolation Kit and other reagents were purchased from Sigma Aldrich. Antibodies raised against DGCR8, β-actin, anti-rabbit and anti-mouse IgG HRP-linked antibody; BCA Protein Assay Kit; RIPA buffer; phosphatase inhibitor cocktail; chemiluminescent western pico plus chemiluminescent substrate, Power up SybrGreen Master Mix, High Capacity Reverse Transcriptions Kit, TaqMan Advanced miRNA cDNA Synthesis Kit and miRNA Advanced Taqman assays were purchased from Thermo Fisher Scientific, Inc. The polyvinylidene difluoride (PVDF) membranes were obtained from Amersham. DICER, DROSHA, XPO5, CTGF, SPARC and GAPDH antibodies were obtained from Santa Cruz Biotechnology. Antibodies raised against phospho-p70 S6K and p70 S6K were purchased from Cell Signaling Technology, Inc. GapmeRs were purchased from Qiagen.
Siddeek B., Mauduit C., Chehade H., Blin G., Liand M., Chindamo M., Benahmed M, & Simeoni U. (2019). Long-term impact of maternal high-fat diet on offspring cardiac health: role of micro-RNA biogenesis. Cell Death Discovery, 5, 71.
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2

Protein Immunoprecipitation and Western Blot

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The following antibodies were used for immuno-precipitation and Western Blot experiments, according to the manufacturer's instruction: DGCR8 (Abcam ab90579), EWSR1 (Abcam ab54708), FUS (Bethyl A300–293A), DDX5 (Abcam ab126730), Drosha (Santa Cruz sc-33778), Vinculin (VCL) (Millipore 06–866), PRMT1 (Abcam ab73246), ASYM24 (Millipore), SYM10 (Millipore), Mono-Methyl Arginine (R*GG) (D5A12) (Cell Signaling Technology 8711), LAMIN A/C (sc-6215), Lamin B1 (Abcam ab16048), GAPDH (Abcam ab9484), H3 (Abcam ab1791), H4 (Abcam ab7311), H4R3me2a (Active Motif 39705), TAF15 (Bethyl Laboratories A300–308A), ILF3 (Bethyl A303–615A), ILF2 (sc-271718), DDX17 (sc-130650), HDAC1 (Abcam ab7028) and HA.11 (Biolegend 901513).
MS023 and MS094 compounds were kindly provided by the SGC Toronto—Structural Genomic Consortium (http://www.thesgc.org/scientists/groups/toronto). Compounds were dissolved in DMSO and used at a final concentration of 10 μM for the indicated time intervals.
Spadotto V., Giambruno R., Massignani E., Mihailovich M., Maniaci M., Patuzzo F., Ghini F., Nicassio F, & Bonaldi T. (2019). PRMT1-mediated methylation of the microprocessor-associated proteins regulates microRNA biogenesis. Nucleic Acids Research, 48(1), 96-115.
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3

Molecular Regulation of miRNA-195 in Cultured Cells

Cited 1 time
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Caco-2 and IEC-6 cells were purchased from the American Type Culture Collection (Manassas, VA) and were maintained in standard culture conditions.18 (link) Antibodies recognizing p53, JunD, HuR, AUF1, DROSHA, and GAPDH were obtained from Santa Cruz Biotechnology (Santa Cruz, CA) and BD Biosciences (Sparks, MD). The secondary antibody conjugated to horseradish peroxidase was obtained from Sigma. Locked nucleic acid (LNA)-modified anti-uc.173 oligonucleotides that antagonize uc.173 and control LNA-scrambled oligonucleotides were custom-generated by Exiqon (Vadbaek, Denmark). Pre-miR™ miRNA precursor of miRNA195 (pre-miR-195) and anti-miR™ inhibitor of miRNA195 (anti-miR-195) were purchased from Ambion (Austin, TX). Biotin-labeled mature and pri-miR-195 were custom-made by Dharmacon (Lafayette, CO).
Xiao L., Wu J., Wang J.Y., Chung H.K., Kalakonda S., Rao J.N., Gorospe M, & Wang J.Y. (2017). Long Noncoding RNA uc.173 Promotes Renewal of the Intestinal Mucosa by Inducing Degradation of MicroRNA 195. Gastroenterology, 154(3), 599-611.
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4

Hippocampus Protein Extraction and Analysis

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The hippocampus proteins were prepared using RIPA buffer (Thermo Fisher Scientific, MA, US), followed by protein quantification using BCA Protein Assay Kit (Thermo Fisher Scientific, MA, US). The western blot was done similarly, as we previously described, using antibodies against ANG (Santa Cruz sc-74528, TX, US), Drosha (Santa Cruz sc-393591, TX, US), Dicer (Cell Signaling Technology #3363, MA, US), or GAPDH (Santa Cruz-47724, TX, US) antibodies [25 (link)].
Wu W., Lee I., Spratt H., Fang X, & Bao X. (2021). tRNA-Derived Fragments in Alzheimer’s Disease: Implications for New Disease Biomarkers and Neuropathological Mechanisms. Journal of Alzheimer's disease : JAD, 79(2), 793-806.
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