To investigate the effect and mechanism of the CXC chemokine receptor 4 (CXCR4) in the proliferation and migration of breast cancer, a short-hairpin RNA (shRNA) eukaryotic expression vector targeting CXCR4 was constructed, and the impact of such on the proliferation, adhesion and migration of human breast cancer MDA-MB-231 cells was observed. The fragments of CXCR4-shRNA were synthesized and cl...

The objectives of the present study were to construct the recombinant primate lentivirus‑short hairpin RNA-pleiotrophin (pLV-shRNA-PTN) vector, to investigate the silencing effect of pLV-shRNA-PTN on PTN expression in MIA PaCa-2 cells and to observe the inhibition efficiency of pLV-shRNA‑PTN on neurite outgrowth from dorsal root ganglion (DRG) neurons in vitro. The construction procedure for re...

To successfully apply RNAi, the very first step is to design functional siRNA or shRNA sequences targeting specific genes. Many characteristics of functional synthetic siRNAs have been identified, but it remains questionable if these characteristics can be equally applied to the design of functional shRNAs. Our three-phase algorithm was developed based on the characteristics of functional siRNA...

RNAi screening using pooled shRNA libraries is a valuable tool for identifying genetic regulators of biological processes. However, for a successful pooled shRNA screen, it is imperative to thoroughly optimize experimental conditions to obtain reproducible data. Here we performed viability screens with a library of ∼10,000 shRNAs at two different fold representations (100- and 500-fold at trans...

To address the unsolved question of PSD-95 function I used short hairpin RNA (shRNA) to knock-down protein expression. shRNA mediated knock-down of protein expression is one of the established methods exploiting the naturally occurring expression control mechanism known as RNA interference (RNAi). Cells express shRNAs that are complementary to the mRNA they control. The shRNAs are processed and...

This study evaluated the effectiveness of a parent based intervention (PBI) in reducing drinking among first year college students (N=443). Students were assigned to one of three conditions: PBI, PBI plus booster brochures (PBI-B), and an assessment only control group (CNT). At a 4-month post-intervention follow-up, results indicated students in the PBI-B group reported significantly less drink...

RNA interference (RNAi) is widely used to determine the function of genes. We chose this approach to assess the collective function of the highly related reproductive homeobox 3 (Rhox3) gene paralogs. Using a Rhox3 short hairpin (sh) RNA with 100% complementarity to all 8 Rhox3 paralogs, expressed from a CRE-regulated transgene, we successfully knocked down Rhox3 expression in male germ cells i...

Duplex RNA harboring the 5'-terminal triphosphate RNA is hypothesized to not only execute selective gene silencing via RNA interference, but also induce type I interferon (IFN) through activation of the retinoic acid inducible gene I (RIG-I). We evaluated gene silencing efficacy of the shRNA containing 5'-triphosphate (3p-shRNA) targeting the hepatitis C virus (HCV) RNA genome in hepatic cells....

BACKGROUND RNA interference (RNAi) technology is a powerful methodology recently developed for the specific knockdown of targeted genes. RNAi is most commonly achieved either transiently by transfection of small interfering (si) RNA oligonucleotides, or stably using short hairpin (sh) RNA expressed from a DNA vector or virus. Much controversy has surrounded the development of rules for the desi...

BioTechniques 625 Adenoviral shuttle vectors to express short hairpin RNAs (shRNA) using RNA polymerase III (RNA pol III) promoters (mouse U6, human H1, or human U6) have been constructed in several laboratories including Welgen (Worcester, MA, USA), BD Biosciences (San Jose, CA, USA), Invitrogen (Carlsbad, CA, USA), and GeneScript (Piscataway, NJ, USA). Welgen and Invitrogen developed adenovir...