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Lentiviral miRNA Expression - Lenti-miR™

Lentiviral miRNA Expression - Lenti-miR™

There are expected to be over 1,000 microRNAs encoded in the human and mouse genomes. While the number of verified human microRNAs is expanding, there is an increasing need for effective functional testing.

Cambridge Bioscience offers the human and mouse Lenti-Mir™ collection from System Biosciences for the stable and robust over expression of miRNA from miRNA precursor clones. These precursor clones can be used to modulate the expression of their targeted miRNAs in vivo. This unique range is the largest and most comprehensive collection of miRNA precursor clones in lentiviral vectors on the market and also is the only commercially-available collection of microRNAs that are cloned in lentiviral vectors capable of being propagated

Benefits of Using the Lenti-miR™ Collection
The Lenti-miR™ clones offer numerous advantages over other miRNA collections:
Largest collection of miRNA precursor clones in HIV-based lentiviral vectors on the market
Only commercially-available collection of microRNAs that are cloned in Lentiviral vectors capable of being propagated
Mouse & human clones available
• Reliable delivery to both dividing & non-dividing cells
Transfection & infection compatible
Stably expressed in multiple cell types
Unusual structure ensure constructs are processed in the cell into mature miRNA
• Pre-microRNA expression from a constitutive CMV promoter
• Enables the monitoring of positive expressing cells with copGFP reporter under the control of the constitutive human elongation factor 1α (EF1) promoter
• Specific clusters available
• Pools of prepackaged viral particles containing the entire microRNA precursor collection for high-throughput phenotypic screens available

Lenti-miR™ Structure
Each of the contructs in the Lenti-miR™ collection are built in a lentiviral vector backbone, so they can be packaged into lentiviral particles and delivered to primary cells, stem cells or other hard-to-transfect cell lines. Therefore unlike other synthetic microRNAs that can only be transiently expressed in cells, these constructs can be stably expressed in many cell types.Unusually, each construct consists of the native stem loop structure and also 200-400 base pairs of upstream and downstream flanking genomic sequence. This unique feature ensure that the miRNAs expressed from these constructs are correctly processed in the cell into mature miRNA.

miRNA Structure
Clusters
The Lenti-Mir™ range includes the following clusters associated with various cancers:
Cancer & Stem Cell miRNA Clusters
• miR-302bcad
• miR-17~92a
• miR-371-371
• miR-106b-25
• miR-106a-363

Breast Cancer Stem Cell Clusters
• miR-200b-200a-429
• miR-200c-141
• miR-183-182

Lenti-miR™ Process

LentiMir Process
To view the Lenti-miR™ collection flyer (PDF), please Click Here

Pooled Pre-packaged miRNA Virus Library- Lenti-miR™
Cambridge Bioscience also offers pools of pre-packaged, ready-to-infect viral particles containing the entire microRNA precursor collection for high-throughput phenotypic screens.

Custom Constructs
Cambridge Bioscience is also able to offer access to a custom construct generation service. To find out more, please Click Here

References
The Lenti-miR constructs have been referenced in numerous publications including:
Hu S, Wilson KD, Ghosh Z, Han L, Wang Y, Lan F, Ransohoff KJ, Burridge P, Wu JC. MicroRNA-302 Increases Reprogramming Efficiency via Repression of NR2F2. Stem Cells. 2013 Feb;31(2):259-68.

Li Y, Gao L, Luo X, Wang L, Gao X, Wang W, Sun J, Dou L, Li J, Xu C, Wang L, Zhou M, Jiang M, Zhou J, Caligiuri MA, Nervi C, Bloomfield CD, Marcucci G, Yu L. Epigenetic silencing of microRNA-193a contributes to leukemogenesis in t(8;21) acute myeloid leukemia by activating the PTEN/PI3K signal pathway. Blood. 2013 Jan 17;121(3):499-509.

Kong W, He L, Richards EJ, Challa S, Xu CX, Permuth-Wey J, Lancaster JM, Coppola D, Sellers TA, Djeu JY, Cheng JQ. Upregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL and is associated with poor prognosis and triple-negative breast cancer. Oncogene. 2013 Jan 28. doi: 10.1038/onc.2012.636. [Epub ahead of print].

Complete Packaging System
For viral transduction we recommend using the pPACK-H1 Lentivector Packaging System (cat.#LV500A-1) together with the 293TN cell line (cat.#LV900A-1). The packaging system provides all the necessary coat proteins to manufacture pseudoviral particles for delivery of the anti-microRNA expression construct into any mammalian cells. In addition the constructs are also suitable for direct transfection into target cells.

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