Synthetic Protein Mimics for Functional Protein Delivery

Area of Science:

• Biotechnology
• Polymer Chemistry
• Cell Biology

Background:

• Protein delivery into cells is hindered by the plasma membrane barrier.
• Existing methods like TAT peptide fusion have limitations.
• Need for efficient, non-covalent protein delivery systems.

Purpose of the Study:

• To develop a novel class of protein transporter molecules.
• To synthesize protein transduction domain mimics (PTDMs) via ring opening metathesis polymerization (ROMP).
• To demonstrate the efficacy of PTDMs in delivering functional proteins into hard-to-transfect cells.

Main Methods:

• Synthesis of PTDMs using ROMP, inspired by amphiphilic peptides.
• Optimization of PTDMs by varying hydrophobic segment length and cationic groups.
• Delivery of Enhanced Green Fluorescent Protein (EGFP) into Jurkat T cells.
• Delivery of Cre Recombinase and Runx1.d190 into human T cells and murine splenocytes, respectively.

Main Results:

• Established the importance of a longer hydrophobic segment with cationic guanidinium groups for efficient delivery.
• Achieved ∼80% knockdown efficiency using Cre Recombinase delivery into human T cells.
• Demonstrated a 2-fold increase in c-Myc mRNA production by delivering Runx1.d190 into murine splenocytes.

Conclusions:

• PTDMs synthesized via ROMP represent a versatile and effective platform for protein delivery.
• This technology overcomes the challenge of protein transport across cell membranes.
• The platform shows potential for therapeutic applications and biological research in hard-to-transfect cells.