Universal Template-Assisted, Cloning-free Method for the Generation of Small RNA-Expressing Dumbbell-Shaped DNA Vectors

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

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 devel...

Small-interfering-RNA expression in cells based on an efficiently constructed dumbbell-shaped DNA.

A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA.

We describe the use of lentiviral vectors expressing small interfering RNAs (siRNAs) to knock down the expression of specific genes in vitro and in vivo. A lentiviral vector capable of generating siRNA specific for GFP after transduction of 293T-GFP cell lines showed no GFP fluorescence. Furthermore, no GFP-specific RNA could be detected. When eggs from GFP-positive transgenic mice were transdu...

Template Assisted Method for Enhancing the Zeolite’s Ability to Capture Methionine

To investigate template effect on physicochemical properties of zeolite and enhance the molecular sieving properties, adsorption of methionine amino acid was studied by spectroscopic methods. Four zeolites such as methionine templated analcime, methionine templated zeolite P, N, N-dibenzyl N,N,N,N-tetramethylethylenediamine (DBTMED) templated analcime and free template analcime were synthesized...

Universal restriction site-free cloning method using chimeric primers.

A universal restriction site-free cloning method has been developed to precisely insert a DNA fragment into a vector at any desired location without altering any nucleotide(s) in either the DNA fragment or the vector. The technique employs two pairs of chimeric primers, each containing a ribonucleotide. One pair of primers is used to amplify a target DNA fragment and another is used to prepare ...