Effective sirna oligonucleotides

Manufactured by RiboBio

Sourced in China

Effective siRNA oligonucleotides are short, double-stranded RNA molecules designed to induce gene silencing by targeting and degrading specific messenger RNA (mRNA) sequences within cells. These oligonucleotides play a core function in RNA interference (RNAi) technology, which is used to regulate gene expression and study gene function.

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Lab products found in correlation

Lipofectamine rnaimax, by Thermo Fisher Scientific (5 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Puromycin, by Merck Group (2 mentions) Sictrl, by RiboBio (1 mentions) Goscript reverse transcription system, by Promega (1 mentions) Sitak1, by RiboBio (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Itaq universal sybr green supermix, by Bio-Rad (1 mentions) Pcdna3.1 vector, by Thermo Fisher Scientific (1 mentions) Penter vector, by Charles River Laboratories (1 mentions) Lv3 lentiviral vector, by GenePharma (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Ex egfp lv105, by GeneCopoeia (1 mentions)

6 protocols using effective sirna oligonucleotides

Targeting TAK1 and NRF2 with siRNA

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Effective siRNA oligonucleotides targeting TAK1 and NRF2 were obtained from RiboBio with the following sequences:
siTAK1-#1: GGAGTGGCTTATCTTCACA;
siTAK1-#2: GGCTTATCTTACACTGGAT;
siNRF2-1#: GAGAAAGAATTGCCTGTAA;
siNRF2-2#: GCTACGTGATGAAGATGGA;
siNRF2-3#: GCCCTCACCTGCTACTTTA.
The negative control (siCtrl) was nonhomologous to any human genome sequence and purchased from RiboBio Co., Ltd. Predetermined cells were grown in 6-well plates for 12 h and then transfected with 20 nM siRNA mixed with 5 μL of Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s instructions. Thirty-six to 48 h after transfection, the cells were harvested for further analysis.
Total RNA was extracted using TRIzol (Life Technology), reverse transcribed into cDNA using the GoScriptTM Reverse Transcription System (Promega) and analyzed by real-time qPCR on a BIO-RAD Real-time PCR machine using iTaq™ Universal SYBR® Green Super mix (Bio–Rad). The results were normalized to β-actin, and relative values were calculated using the 2^{[-(CT gene -CT reference)]} method. The gene-specific primers used for qPCR are listed in Supplementary Table 3.

Yuan L., Li S., Chen Q., Xia T., Luo D., Li L., Liu S., Guo S., Liu L., Du C., Jia G., Li X., Lu Z., Yang Z., Liu H., Mai H, & Tang L. (2022). EBV infection-induced GPX4 promotes chemoresistance and tumor progression in nasopharyngeal carcinoma. Cell Death and Differentiation, 29(8), 1513-1527.

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Transfection of miR-637 and circHIPK3 in Osteosarcoma

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miR-637 mimics and negative control (NC mimic), miR-637 inhibitors and negative control (NC inhibitor), effective siRNA oligonucleotides that targeted the splicing sites of circHIPK3 (designed using circPrimer; version 1.2.0.5) (29 (link)) or targeted HDAC4 and scrambled control siRNAs were synthesized by Guangzhou RiboBio Co., Ltd. When the OS cells reached 80% confluence, the oligonucleotides (100 nM) were transfected using Lipofectamine^® RNAiMax (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. After 48 h of transfection, the OS cells were used for further detection. The sequences of the oligonucleotides are listed in Table I.

Wen Y., Li B., He M., Teng S., Sun Y, & Wang G. (2020). circHIPK3 promotes proliferation and migration and invasion via regulation of miR-637/HDAC4 signaling in osteosarcoma cells. Oncology Reports, 45(1), 169-179.

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Oligonucleotide Transfection for Lung Cancer

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Effective siRNA oligonucleotides that targeted linc00968 or CPEB3 as well as negative control siRNAs (si-Ctrl), miR-9-5p mimic and negative control (miR-Ctrl mimic), FAM labeled miR-9-5p inhibitor (anti-miR-9-5p) and negative control (anti-miR-Ctrl) were purchased from RiboBio Co., Ltd (Ribobio, Guangzhou, China). All the oligonucleotides and plasmids were transfected into A549, H1975 or HCC827 cell using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The full-length of linc00968 was synthesized by RiboBio (Guangzhou, China) and subcloned into the pcDNA3.1 (+) vector (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer's instructions.

Tang H., Han X., Feng Y, & Hao Y. (2020). linc00968 inhibits the tumorigenesis and metastasis of lung adenocarcinoma via serving as a ceRNA against miR-9-5p and increasing CPEB3. Aging (Albany NY), 12(22), 22582-22598.

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Knockdown of DANCR in NPC Cells

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Effective siRNA oligonucleotides that targeted DANCR, NF45 or NF90 were purchased from RiboBio (Guangzhou, China). LV3 lentiviral vectors that encode short hairpin RNA (shRNA) targeting DANCR or non-specific control were constructed by GenePharma (Suzhou, China). The siRNA and shRNA sequences are shown in Table S1. The pENTER-vector, pENTER-DANCR and pENTER-HIF-1α plasmids were purchased from Vigene Biosciences (Jinan, China). All of the plasmids were confirmed through DNA sequencing.
The synthesized short hairpin RNA targeting DANCR (shDANCR) or control shRNA were cloned into pSuper-retro-puromycin vectors (Addgene, Cambridge, MA, USA). NPC cells were transfected with oligonucleotides (100 nM) or plasmids (2 μg) using Lipofectamine 3000 or Lipofectamine 2000 reagents (Invitrogen). The cells were harvested for assays 48 h after transfection. SUNE-1 cell lines stably expressing shDANCR or vector were generated after lentiviral infection by 293FT cells and selected by 0.5 μg/mL puromycin.

Wen X., Liu X., Mao Y.P., Yang X.J., Wang Y.Q., Zhang P.P., Lei Y., Hong X.H., He Q.M., Ma J., Liu N, & Li Y.Q. (2018). Long non-coding RNA DANCR stabilizes HIF-1α and promotes metastasis by interacting with NF90/NF45 complex in nasopharyngeal carcinoma. Theranostics, 8(20), 5676-5689.

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Knockdown and Overexpression of CRLF1 and MYH9 in PTC

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We bought the effective siRNA oligonucleotides targeting the CRLF1 from Guangzhou Ribobio Company (Guangzhou, China) and transfected using Lipofectamine RNAiMax (Invitrogen) according to the manufacturer's protocol. We purchased the lentiviral vector encoding FLAG-tagged CRLF1 (EX-N0027-Lv121), His-tagged MYH9 (EX-T1335-Lv128), the control vector (EX-EGFP-Lv105), and the packaging system (HIV) from GeneCopeia (USA). Subsequently, we verified all the plasmids via DNA sequencing. The siRNA sequences used are shown in Table S3. We co-transfected the lentivirus packaging expression plasmids into the 293T cells. Then, we collected the supernatants containing viruses and used them to infect the PTC cell lines for 48 h. After that, we selected the stable clones of these cells using puromycin (Sigma-Aldrich, USA) for 7 days after infection. We used qRT-PCR and blotting assays to assay for the expression levels of CRLF1 and MYH9 western.

Yu S.T., Sun B.H., Ge J.N., Shi J.L., Zhu M.S., Wei Z.G., Li T.T., Zhang Z.C., Chen W.S, & Lei S.T. (2020). CRLF1–MYH9 Interaction Regulates Proliferation and Metastasis of Papillary Thyroid Carcinoma Through the ERK/ETV4 Axis. Frontiers in Endocrinology, 11, 535.

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Silencing CRLF1 in Thyroid Cancer

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Effective siRNA oligonucleotides that targeting CRLF1 were purchased from Guangzhou Ribobio Company (Guangzhou, China) and were transfected using Lipofectamine RNAiMax (Invitrogen) according to the manufacturer’s instructions. The lentiviral vector encoding FLAG-tagged CRLF1 (EX-N0027-Lv121), the control vector (EX-EGFP-Lv105), and the packaging system (HIV) were obtained from GeneCopeia (USA). All the plasmids were verified by DNA sequencing. The siRNA sequences used were as follows:
siRNA 1# of CRLF1 sense 5′-GGCUCUCUUACGCCCUAU dTdT-3′;
anti-sense 5′-AUAGGGCGUAAAGAGAGCC dTdT-3′;
siRNA 2# of CRLF1 sense 5′-CACGCUGGAUAUCCUGGAU dTdT-3′;
anti-sense 5′-GUGCGACCUAUAGGACCUA dTdT-3′.
The lentivirus packaging expression plasmids were co-transfected into 293T cells. Then, the supernatants containing viruses were collected and used to infect the PTC cell lines for 48 h. Then, stable clones of these cells were selected with puromycin (Sigma-Aldrich, USA) for 7 days after infection. The expression levels of CRLF1 were verified by qRT-PCR and western blotting assays.

Yu S.T., Zhong Q., Chen R.H., Han P., Li S.B., Zhang H., Yuan L., Xia T.L., Zeng M.S, & Huang X.M. (2018). CRLF1 promotes malignant phenotypes of papillary thyroid carcinoma by activating the MAPK/ERK and PI3K/AKT pathways. Cell Death & Disease, 9(3), 371.

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