Efficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression.

ABSTRACT: Genetic therapy holds great promise for the treatment of inherited or acquired genetic diseases; however, its breakthrough is hampered by the lack of suitable gene delivery systems. Dumbbell-shaped DNA minimal vectors represent an attractive, safe alternative to the commonly used viral vectors which are fraught with risk, but dumbbell generation appears to be costly and time-consuming. We developed a new PCR-based method for dumbbell production which comprises only two steps. First, PCR amplification of the therapeutic expression cassette using chemically modified primers to form a ready-to-ligate DNA structure; and second, a highly efficient intramolecular ligation reaction. Compared with conventional strategies, the new method produces dumbbell vectors more rapidly, with higher yields an

SUBMITTER: Yu H 

PROVIDER: S-EPMC4605290 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature