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EXP001866

Paper

Delivery of therapeutic shRNA and siRNA by Tat fusion peptide targeting bcr–abl fusion gene in Chronic Myeloid Leukemia cells (2010)

Peptide

Tat–LK15

Sequence: RKKRRQRRRGGGKLLKLLLKLLLKLLK-CONH2

RNA

siRNA

All experiment fields

Experiment Id EXP001866
Paper Delivery of therapeutic shRNA and siRNA by Tat fusion peptide targeting bcr–abl fusion gene in Chron
Peptide Tat–LK15
Delivery Success Class no
In Vivo Flag no
Uptake Confirmed yes
Label Confidence high
In Vitro Functional Effect yes
Endosomal Escape Evidence
Peptide Concentration Example prep (1:1 charge ratio): 0.81 µg peptide in 50 µL HEPES-buffered saline mixed with 1 µg oligo in 50 µL; incubate 30 min RT. Higher charge ratios used for functional assays.
Rna Concentration siRNA 1–30 µg (≈60 nM–1.8 µM range) tested; key readout at 48 h post-transfection
Mixing Ratio (+/−) charge ratio 3:1 (Tat–LK15:siRNA) for silencing studies
Formulation Format Noncovalent electrostatic peptide/nucleic acid complexes
Formulation Components Tat–LK15 fusion peptide complexed with BCR–ABL-targeting siRNA or shRNA expression plasmid (pSilencer 2.0 U6 hygro)
Size Nm 663.40
Zeta Mv 10.55
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells K562 human chronic myelogenous leukemia cells
Animal Model
Administration Route
Output Type Gene silencing (BCR–ABL protein by Western blot at 48 h)
Output Value ≥~70% BCR–ABL protein reduction at 48 h across wide siRNA amounts (max around 5–10 µg); silencing not observed beyond 48 h (transient)
Output Units
Output Notes Cy5-siRNA uptake by FACS up to ~60% positive cells; higher charge ratio associated with higher toxicity vs shRNA complexes.
Toxicity Notes MTT: siRNA complexes significantly more toxic than shRNA complexes at 24 h; Tat–LK15 alone high toxicity at higher concentrations.
Curation Notes