Let-7e-5p-GFP-Puro

Background

Let-7e is a member of the highly conserved let-7 family of microRNAs, among the first microRNAs identified and central regulators of developmental timing and cell differentiation. Like other microRNAs, let-7e post-transcriptionally silences target mRNAs by guiding the RNA-induced silencing complex to complementary sequences, causing translational repression or transcript decay. The let-7 family acts broadly as tumor-suppressive microRNAs, restraining proliferation by repressing oncogenic targets such as RAS and MYC, and its expression is frequently reduced in cancer. Let-7e thus contributes to control of cell growth, differentiation, and tissue homeostasis and is widely studied in cancer biology.

Product Description & Applications

The Let-7e-5p-GFP-Puro Lentivirus delivers stable overexpression of human and mouse let-7e. The vector carries the entire miRNA stem-loop, allowing endogenous Drosha and Dicer processing to generate the authentic mature let-7e-5p strand for loading into the RNA-induced silencing complex. The stem-loop is driven by a U6 promoter, while a PGK promoter drives a GFP fluorescent reporter and a puromycin selection marker separated by a self-cleaving peptide. Transduced cells are readily identified and enriched by GFP fluorescence or puromycin selection, with polyclonal stable lines established within about ten days while preserving parental cell properties. It is used for long-term target-repression studies, tumor models, and other functional microRNA experiments. Supplied as ultra-purified, high-titer lentiviral particles suitable for in vitro and in vivo use and for primary or thawed cells.

About This Product

This lentivirus expresses the full pre-miRNA stem-loop sequence of MIRLET7E under a Pol II promoter (CMV or EF1α), enabling endogenous Drosha/DGCR8 and Dicer processing to produce authentic mature 5P (and 5p) strands loaded into RISC. This produces physiologically relevant miRNA activity that faithfully replicates the endogenous biogenesis pathway, unlike synthetic miRNA mimics that bypass nuclear processing and can exhibit RISC overloading artifacts.

Co-expression of a fluorescent reporter (GFP) and selection marker (Puromycin) allows stable transductant identification by fluorescence and antibiotic enrichment. Stable miRNA overexpression enables long-term target repression studies, in vivo tumor models, and exosomal miRNA loading experiments that are not achievable with transient miRNA mimics.