TY - JOUR AU - Frank, Sander B. AU - Schulz, Veronique V. AU - Miranti, Cindy K. PY - 2017 DA - 2017/02/28 TI - A streamlined method for the design and cloning of shRNAs into an optimized Dox-inducible lentiviral vector JO - BMC Biotechnology SP - 24 VL - 17 IS - 1 AB - Short hairpin RNA (shRNA) is an established and effective tool for stable knock down of gene expression. Lentiviral vectors can be used to deliver shRNAs, thereby providing the ability to infect most mammalian cell types with high efficiency, regardless of proliferation state. Furthermore, the use of inducible promoters to drive shRNA expression allows for more thorough investigations into the specific timing of gene function in a variety of cellular processes. Moreover, inducible knockdown allows the investigation of genes that would be lethal or otherwise poorly tolerated if constitutively knocked down. Lentiviral inducible shRNA vectors are readily available, but unfortunately the process of cloning, screening, and testing shRNAs can be time-consuming and expensive. Therefore, we sought to refine a popular vector (Tet-pLKO-Puro) and streamline the cloning process with efficient protocols so that researchers can more efficiently utilize this powerful tool. SN - 1472-6750 UR - https://doi.org/10.1186/s12896-017-0341-x DO - 10.1186/s12896-017-0341-x ID - Frank2017 ER -