本文采用的英格恩产品: Entranster-H4000

Rapidly Visualizing the Membrane Affinity of Gene Vectors Using Polydiacetylene-Based Allochroic Vesicles

Jing-Wen Wang  ¹ , Feng Zheng  ¹ , Huan Chen  ² , Ya Ding  ¹ , Xing-Hua Xia  ³ Affiliations

• PMID: 30896923
• DOI: 10.1021/acssensors.9b00102

Abstract

The high-throughput screening of chemically active substances has aroused widespread interest in recent years. The screening of drug carriers is usually ignored, although they interact directly with physiological barriers and target cells, and they determine the fate and efficacy of drugs in vivo. In this work, a series of polydiacetylene (PDA) vesicles (ca. 550 nm) that simulate the cell membrane are constructed to detect the membrane affinity of gene vectors. The surface potentials of vesicles are adjusted by changing the phospholipid composition using different charged compounds. All vesicles show the rapid color changes upon the addition of gene vectors by the naked eye within <5 min. The optimized 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC)-PDA vesicles display the most sensitive discoloration response to the commercially available gene vectors, including Lipofectamine 2000, Entranster-H4000, and polyethylenimine. The logarithm of transfection efficiency for green fluorescent protein plasmid (pGFP) mediated by these three vectors in L02 and HepG2 cells demonstrate an excellent linear correlation with the logarithm of membrane affinity (log K_b) of the gene vectors detected by DMPC-PDA vesicles. This rapid visualization method not only allows the in vitro membrane affinity prediction of gene vectors that greatly contributes to the gene transfection efficiency, but also offers a universal strategy for the potential high-throughput screening of various carrier materials featuring high cell affinity.

Keywords: allochroic polydiacetylene vesicles; gene vector; high-throughput screening; membrane affinity; transfection efficiency.