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13. The cell penetrating peptides TAT and antennapedia can deliver functional biological cargo in vitro

We examined a number of cell penetrating peptides (CPPs) for their ability to deliver functional biological cargo (short peptides and siRNA anti-sense) in vitro in a range of different cell types. In addition, in order to assess the suitability of different CPPs for biological studies we also looked at cellular toxicity.

In all we examined the following CPPs -TAT, D-TAT, TAT(+ K), antennapedia, transportan and polyarginine. With the attachment of a short peptide cargo we examined both the uptake and cytotoxicity in HeLa, A549 and CHO K1 cells. Based on uptake and cytotoxicity our results showed that both TAT and antennapedia were the "best" CPPs for peptide delivery and that rate and amount of uptake was largely independent of cell type.

Using TAT and antennapedia CPPs we were also able to deliver functional siRNA anti-sense material both in vitro and in vivo. Firstly, in vitro using TAT-siRNA conjugates directed against p38 MAPK we achieved 80% and 50% knockdown in human and mouse cell lines respectively. Both TAT and antennapedia siRNA conjugates also showed functional activity in vivo, lowering the endogenous level of p38 in the mouse lung by 40% following intra-tracheal administration.

The demonstration of the application of CPPs for the cellular delivery of biological cargo is potentially of great interest for the future utility of alternative non-viral delivery systems both for in vitro and in vivo target validation studies and future therapeutics.

Contact

Simon JONES, (Post-doctoral Research Scientist)
Tel.: +1-625-517340
E-mail
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