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Klf4 sirna

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KLF4 siRNA is a targeted small interfering RNA (siRNA) designed to silence the expression of the Krüppel-like factor 4 (KLF4) gene. It is a molecular research tool used to study the role of KLF4 in cellular processes.

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Lab products found in correlation

Scrambled sirna, by Thermo Fisher Scientific (3 mentions) Hylorondase, by Thermo Fisher Scientific (2 mentions) Hor 272, by Merck Group (2 mentions) Klf5 sirnas, by Thermo Fisher Scientific (2 mentions) Human chorionic gonadotropin (hcg), by Merck Group (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions) Scramble control, by OriGene (1 mentions) Pmir report vector, by Thermo Fisher Scientific (1 mentions) Siport neofx transfection agent, by Thermo Fisher Scientific (1 mentions) Antagomir 200a, by RiboBio (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Control nontargeting scrambled sirna duplexes, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

5 protocols using klf4 sirna

1

KLF4 Silencing in Muse Cells

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Three kinds of Klf4 siRNA (Ambion, Thermo Fisher Scientific, ID: S17793, S17794, and S17795) were mixed up, and naive Muse cells were sorted and transfected with 25 pmol Klf4 siRNA or 25 pmol scrambled siRNA (Ambion) by Lipofectamine RNAiMAX (Invitrogen, 13778–150) in 6-well plates for 48 h following the manufacturer’s instructions. Total RNA was extracted, and gene expression was analyzed by qPCR as described above.
Li G., Wakao S., Kitada M, & Dezawa M. (2024). Tumor suppressor let-7 acts as a key regulator for pluripotency gene expression in Muse cells. Cellular and Molecular Life Sciences: CMLS, 81(1), 54.
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2

Keap1 3'-UTR Regulation by miR-200a

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pGL3-ARE and pcDNA3-HA-Nrf2 were generous gifts from Professor Xiaoming Yang. The 3′-UTR of Keap1 was amplified using the following primers: 5′-TCATACTAGTGGCACTTTTGTTTCTTGGGC-3′ and 5′-GCATTAAGCTTCAGGGTGAAAGACACTAG-3′ and cloned into pMiR-Report vector (Ambion) digested with HindIII and Spe I. We also generated three bases mutation in the predicated target site for miR-200a by using a QuickChange site-specific mutagenesis kit (Stratagene). All constructs were sequenced in Sangon Company. Transfection of plasmids was performed in 70%–80% confluent cells using Lipofectamine 2000 Reagent (Invitrogen) according to manufacturer's protocol.
MSA was purchased from Sigma–Aldrich (Sigma–Aldrich Inc.). Pre-miR miR-200a precursor and Pre-miR negative control were purchased from Ambion. Antagomir-200a was synthesized from Ribobio. KLF4 siRNA and scramble control were purchased from OriGene (OriGene Technologies). Transfections of Pre-miR miR-200a precursor, Pre-miR negative control, KLF4 siRNA and scramble control were performed by using siPORT NeoFX Transfection Agent (Life Technologies) according to manufacturer's protocol.
Liu M., Hu C., Xu Q., Chen L., Ma K., Xu N, & Zhu H. (2015). Methylseleninic acid activates Keap1/Nrf2 pathway via up-regulating miR-200a in human oesophageal squamous cell carcinoma cells. Bioscience Reports, 35(5), e00256.
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3

Silencing Transcription Factors in Keratinocytes

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FOXC1 siRNA duplexes, ZNF750 siRNA duplexes, KLF4 siRNA and control non-targeting scrambled siRNA duplexes were purchased from Life Technologies. The sequence for KLF4 siRNA are: sense: UGACCAGGCACUACCGUAAtt; antisense: UUACGGUAGUGCCUGGUCAtt. The sequence for ZNF750 siRNA are: sense: CCUCAAUGUUGUGAACGGAtt; antisense: UCCGUUCACAACAUUGAGGct. The sequence for FOXC1 siRNA are: sense 1: GAAAGUCGCUUUCUUUUUAtt; antisense: UAAAAAGAAAGCGACUUUCat. Sense 2: ACUCUCCAGUGAACGGGAAtt, antisense: UUCCCGUUCACUGGAGAGUtg; Sense 3: AGAGGAUCGGCUUGAACAAtt; anti-sense: UUGUUCAAGCCGAUCCUCUgt. KCs were plated in 24 well plates at 1x105 per well the day before transfection. Cells were transfected with siRNA duplexes at final concentration of 10 nM using lipofectamine 2000 according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA). After 24 hours incubation, the cells were replaced with EpiLife supplemented either with 0.06mM or 1.3 mM calcium for up to 120 hours. Then the cells were used for total RNA and protein extraction.
Bin L., Deng L., Yang H., Zhu L., Wang X., Edwards M.G., Richers B, & Leung D.Y. (2016). Forkhead Box C1 Regulates Human Primary Keratinocyte Terminal Differentiation. PLoS ONE, 11(12), e0167392.
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4

Embryo Manipulation via siRNA Knockdown

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5IU of PMSG (Prospec Cat# HOR-272) and HCG (Sigma-Aldrich, Cat # CG5–1VL) were injected intraperitoneally into 4–5 week old, wild-type C57BL/6J female or or C57BL/6J C3H/HeJ F1 mice 46–48h apart. Immediately following HCG injection, females were paired with C57BL/6J stud males and pronuclear stage embryos together with cumulus cell clusters were harvested from plugged females 20h after the mating. Pronuclear stage embryos were dissociated from cumulus cells using Hylorondase (Fisher, Cat # MR-0510F). Embryos were then incubated at 37C with 5% CO2 while siRNAs were prepared for injection. Before injection, scrambled siRNA (Thermo Fisher Scientific, Cat# AM4611) and Klf5 siRNAs (Thermo Fisher Scientific, Cat# 160900) and Klf4 siRNAs (Thermo Fisher Scientific, Cat# 156021) were prepared at a working concentration of 100uM were spun at 10k rpm for 5 min to clear debris. In experiments when we double knocked down Klf4 and Klf5, we prepared siRNA mixtures containing 50 uM Klf4 siRNAs and 50 uM Klf5 siRNAs. After the microinjection of siRNAs, embryos were cultured in vitro to E4.0, and then subjected to IF analyses described above.
Kinisu M., Choi Y.J., Cattoglio C., Liu K., de Bezieux H.R., Valbuena R., Pum N., Dudoit S., Huang H., Xuan Z., Kim S.Y, & He L. (2021). Klf5 establishes bi-potential cell fate by dual regulation of ICM and TE specification genes. Cell reports, 37(6), 109982.
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5

Embryo Manipulation via siRNA Knockdown

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5IU of PMSG (Prospec Cat# HOR-272) and HCG (Sigma-Aldrich, Cat # CG5–1VL) were injected intraperitoneally into 4–5 week old, wild-type C57BL/6J female or or C57BL/6J C3H/HeJ F1 mice 46–48h apart. Immediately following HCG injection, females were paired with C57BL/6J stud males and pronuclear stage embryos together with cumulus cell clusters were harvested from plugged females 20h after the mating. Pronuclear stage embryos were dissociated from cumulus cells using Hylorondase (Fisher, Cat # MR-0510F). Embryos were then incubated at 37C with 5% CO2 while siRNAs were prepared for injection. Before injection, scrambled siRNA (Thermo Fisher Scientific, Cat# AM4611) and Klf5 siRNAs (Thermo Fisher Scientific, Cat# 160900) and Klf4 siRNAs (Thermo Fisher Scientific, Cat# 156021) were prepared at a working concentration of 100uM were spun at 10k rpm for 5 min to clear debris. In experiments when we double knocked down Klf4 and Klf5, we prepared siRNA mixtures containing 50 uM Klf4 siRNAs and 50 uM Klf5 siRNAs. After the microinjection of siRNAs, embryos were cultured in vitro to E4.0, and then subjected to IF analyses described above.
Kinisu M., Choi Y.J., Cattoglio C., Liu K., de Bezieux H.R., Valbuena R., Pum N., Dudoit S., Huang H., Xuan Z., Kim S.Y, & He L. (2021). Klf5 establishes bi-potential cell fate by dual regulation of ICM and TE specification genes. Cell reports, 37(6), 109982.
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