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 Bioengjournal_title
Biotechnology and bioengineeringauthors
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 Fdoi
10.1002/bit.27245subject
Has Abstractpub_date
2020-03-01 00:00:00pages
816-831issue
3eissn
0006-3592issn
1097-0290journal_volume
117pub_type
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