✓Design and express cell-permeable fusion proteins using protein transduction domains.
✓Purify recombinant proteins to homogeneity for cellular delivery experiments.
✓Deliver biologically active proteins directly into cells without genetic modification.
✓Evaluate protein transduction efficiency and cellular outcomes quantitatively.
Protocol
Protein transduction enables the direct delivery of biologically active proteins into cells. In contrast to conventional methods such as DNA transfection or viral transduction this non-invasive paradigm allows highly efficient cellular manipulation in a titratable manner circumventing cellular toxicity and the risk of oncogenic transformation by permanent genetic modification.
Introduce protein transduction paradigm and rationale
Overview of protein transduction as a non-invasive alternative to DNA transfection and viral vectors, emphasizing titratable delivery and avoidance of genomic toxicity.
▶ 01:38
2
Express cell-permeable fusion protein in cells
Culture and induce mammalian or bacterial expression systems to produce recombinant proteins fused to cell-penetrating peptide domains.
▶ 02:34
3
Purify fusion protein to homogeneity
Apply chromatography and biochemical purification techniques to isolate and concentrate the engineered protein from cell lysates or culture medium.
▶ 06:54
4
Perform protein transduction into target cells
Incubate purified cell-permeable protein with cultured mammalian cells and quantify intracellular protein accumulation and biological activity.
▶ 13:46
5
Analyze transduction efficiency and functional outcomes
Evaluate protein localization, cellular response, and comparative efficacy relative to conventional delivery methods using microscopy or biochemical assays.
▶ 20:02
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