Figure 1.

AMPM design and construction of AMPM cassettes. A. Nucleotide sequence and predicted secondary structure of AMPMs targeting the GL3 firefly luciferase (AMPM-FL), human p53 (AMPM-p53) and human lamin A/C (AMPM-lamin A/C) genes. The secondary structures of AMPMs were predicted using mfold program (see Materials and Methods). Each AMPM is composed of a 14 nt loop, a stem region containing a mismatch and a bulge structure, and an extended stem region containing two mismatches. The antisense regions indicated as red letters are fully complementary to the target sequences. The modified sense regions are indicated as blue letters. B. The schematic representation of the construction of the AMPM cassette. The multiple cloning sites of the parental vector, pBS-miR, are shown at the top. The oligonucleotide fragments containing the antisense region (red letters) and the modified sense region (blue letters) of AMPM were inserted into pBS-miR in two steps to generate the pBS-AMPM vectors.