Back to browse

EXP001865

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

shRNA plasmid (DNA vector expressing shRNA)

All experiment fields

Experiment Id EXP001865
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 shRNA plasmid 10–30 µg (within total DNA 30 µg); 4 h transfection then culture 96–192 h
Mixing Ratio (+/−) charge ratio 2:1 (Tat–LK15:plasmid DNA) 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 850.00
Zeta Mv 4.09
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; mRNA by qRT-PCR) over time
Output Value Up to ~90% BCR–ABL protein reduction at 96 h (30 µg shRNA plasmid); ~85% reduction still at 192 h with highest shRNA dose; mRNA reduced ~70–85% at 48 h
Output Units
Output Notes Compared vs Lipofectamine controls; negative control shRNA shows no effect.
Toxicity Notes MTT: shRNA plasmid complexes show no significant cytotoxicity at 24 h; peptide alone becomes toxic >10 µM.
Curation Notes