Cell Line Selection for Target Validation

siRNA Transient Gene Knockdown

Knock down target gene by siRNA pool or individual oligos

  • Taqman or Western blot analysis to confirm the knockdown efficiency at mRNA level or protein level, respectively
  • The effect of tumor cell growth inhibition is monitored by CellTiter Glo cell viability assay (2D) and/or soft agar colony formation assay (3D)
Taqman KD Evaluation

Western Blot KD Evaluation (GAPDH as an example)
siRNA Anti-Proliferation CTG Assay in Cell Line X
siRNA Soft Agar Screen in Cell Line X
shRNA-Mediated Gene Knockdown
  • pTRIPZ System (Open Biosystems) – tet-inducible target gene knockdown
  • pSLIK System (ATCC) – tet-inducible target gene knockdown
  • pGIPZ System (Open Biosystems) – constitutive target gene knockdown
  • pLKO1 System (Open Biosystems) – constitutive target gene knockdown


Verify Inducible Target KD in Stable Cell Lines by RT-Taqman

Verify Target KD in Stable Cell Lines by Western blot

Evaluate Growth Inhibition by Cionogenic Assay

Detection of Target A Knockdown and Down-regulation of Methyl Marker in Dox-induced Stable Cell Lines

Stable Cell Line Generation with Lentivirus
  • pTRIPZ System (Open Biosystems) – tet-inducible target gene knockdown
  • pSLIK System (ATCC) – tet-inducible target gene knockdown
  • pGIPZ System (Open Biosystems) – constitutive target gene knockdown
  • pLKO1 System (Open Biosystems) – constitutive target gene knockdown


Verify Inducible Target KD in Stable Cell Lines by RT-Taqman

Verify Target KD in Stable Cell Lines by Western blot

Evaluate Growth Inhibition by Cionogenic Assay

Detection of Target A Knockdown and Down-regulation of Methyl Marker in Dox-induced Stable Cell Lines