High-efficiency protein delivery into transfection-recalcitrant cell types.

Abstract:

:Intracellular delivery of functional proteins is of great interest for basic biological research as well as for clinical applications. Transfection is the most commonly used method, however, it is not applicable to large-scale manipulation and inefficient in important cell types implicated in biomedical applications, such as epithelial, immune and pluripotent stem cells. In this study, we explored a bacterial type III secretion system (Bac-T3SS)-mediated proteofection method to overcome these limitations. An attenuated Pseudomonas aeruginosa vector was constructed, which has features of low toxicity, high T3SS activity, and self-limiting growth. Compared to the method of transfection, the Bac-T3SS showed significantly higher efficiencies of Cre recombinase translocation and target site recombination for hard-to-transfect human cell lines. Furthermore, through the delivery of β-lactamase in live animals, we demonstrated the feasibility and biosafety of in vivo application of the Bac-T3SS. This study provided an efficient and low-cost proteofection strategy for laboratory use as well as for application in large-scale cell manipulations.

journal_name

Biotechnol Bioeng

authors

Li Z,Cai Z,Fu W,Liu Y,Tian C,Wang H,Fu T,Wu Z,Wu D,Jin Y,Cheng Z,Terada N,Liu L,Wu W,Jin S,Bai F

doi

10.1002/bit.27245

subject

Has Abstract

pub_date

2020-03-01 00:00:00

pages

816-831

issue

3

eissn

0006-3592

issn

1097-0290

journal_volume

117

pub_type

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