Product Image

Contributor Information

  • Name Matthias Hentze
  • Institute European Molecular Biology Laboratory (EMBL)

Tool Details

  • Tool name: HeLa Flp-In SLC25A6+SLC7A5 3′ UTR
  • Tool type: Cell Lines
  • Parental cell line: HeLa FLP-IN T-REx
  • Model: Knock-Out
  • Application: YBX3
  • Description: To identify genes whose expression is controlled by YBX3
  • Research area: Metabolism
  • Production details: Adherent HeLa cells were grown at 37°C and 5% CO2 in DMEM, supplemented with 10% FBS Gold (PAA), 1% Penicillin/Streptomycin (Sigma) and 1% L-glutamine (GIBCO). Cells were grown until 70%–80% confluent, trypsinized, counted (Biorad TC20) and 200,000 cells were seeded per well in a 6-well dish for reverse transfection of siRNAs following the manufacturer’s recommendations (Lipofectamine RNAiMax, Invitrogen, cat# 137780755). YBX3 and control siRNAs were obtained from Dharmacon (YBX3 pool: SMARTpool ON-Targetplus L-015793-00 or YBX3 3′ UTR targeting: ON-Targetplus L-015793-07; and control pool: ON-Targetplus non-targeting siRNAs 1-4). The siRNA used to target the YBX3s 3′ UTR was YBX3 smartpool J-015793-07. Cells were harvested 48 hours after transfections.

  • For Research Use Only

Target Details

  • Target: YBX3

Application Details

  • Application: YBX3

Handling

  • Format: Frozen
  • Growth medium: Adherent HeLa cells were grown at 37°C and 5% CO2 in DMEM, supplemented with 10% FBS Gold (PAA), 1% Penicillin/Streptomycin (Sigma) and 1% L-glutamine (GIBCO) Grown in medium containing blasticidine (5mg/ml) and hygromycin (200mg/ml). Cells were grown until 70%–80% confluent, trypsinized, counted (Biorad TC20) and 200,000 cells were seeded per well in a 6-well dish for reverse transfection of siRNAs following the manufacturer’s recommendations (Lipofectamine RNAiMax, Invitrogen, cat# 137780755). YBX3 and control siRNAs were obtained from Dharmacon (YBX3 pool: SMARTpool ON-Targetplus L-015793-00 or YBX3 3′ UTR targeting: ON-Targetplus L-015793-07; and control pool: ON-Targetplus non-targeting siRNAs 1-4). The siRNA used to target the YBX3s 3′ UTR was YBX3 smartpool J-015793-07. Cells were harvested 48 hours after transfections.

Documentation

  • Available on request

References

  •   The RNA-Binding Protein YBX3 Controls Amino Acid Levels by Regulating SLC mRNA Abundance