Enhancement and optimization of plasmid expression in femtosecond optical transfection

Bavishna B. Praveen, David J. Stevenson, Maciej Antkowiak, Kishan Dholakia, Frank J. Gunn-Moore

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

Cell transfection using femtosecond lasers is gaining importance for its proven ability to achieve selective transfection in a sterile and relatively non-invasive manner. However, the net efficiency of this technique is limited due to a number of factors that ultimately makes it difficult to be used as a viable and widely used technique. We report here a method to achieve significant enhancement in the efficiency of femtosecond optical transfection. The transfection procedure is modified by incorporating a suitable synthetic peptide containing nuclear localization and DNA binding sequences, assisting DNA import into the nucleus. We achieved a 3-fold enhancement in the transfection efficiency for adherent Chinese Hamster Ovary (CHO-K1) cells with this modified protocol. Further, in the presence of this biochemical reagent, we were able to reduce the required plasmid concentration by similar to 70% without compromising the transfection efficiency. Also, we report for the first time the successful photo-transfection of recently trypsinised cells with significantly high transfection efficiency when transfected with modified plasmid. This paves the way for the development of high throughput microfluidic optical transfection devices.

Original languageEnglish
Pages (from-to)229-235
Number of pages7
JournalJournal of Biophotonics
Volume4
Issue number4
DOIs
Publication statusPublished - Apr 2011

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