Pooled MicroRNA Library

Screen all microRNAs simultaneously for unique phenotypes

The Lenti-miR™ pooled virus library is a tool that enables the study of phenotypic effects associated with the over-expression of individual microRNAs. The lentivirus preparation is pseudotyped with VSV-G that allows for broad cellular tropism. Hard-to-transfect mammalian cell lines, primary cells, non-dividing cells and even whole animal studies are possible. Transduced cells exhibiting the phenotypes of interest are isolated by selection or sorting. The microRNA or microRNAs responsible for generating the phenotypes of interest may be recovered through simple genomic PCR using lentivector-specific primers followed by direct sequencing of microRNA precursor clones.

Features and Benefits

Perform one transduction and easily identify the microRNAs involved in your phenotypic screen

The Lenti-miR™ virus library contains a pool of SBI’s microRNA precursor clones in a ready-to-infect lentiviral preparation. Each virus within the pool will express an individual miRNA precursor in its native context while preserving putative hairpin structures to ensure biologically relevant interactions with endogenous processing machinery and regulatory partners. Details of the individual microRNA precursor clones and the list of the microRNAs which comprise the virus pool can be found on the Lenti-miR collection.

The microRNA or microRNAs responsible for generating the phenotypes of interest may be recovered through simple genomic PCR using the included lentivector-specific primers followed by direct sequencing. All microRNA precursor clones are about 500-700 bp in size.

Quality Control of Lenti-miR Virus Library

Infectious titer is determined through real-time qPCR using SBI’s UltraRapid Lentiviral Titer Kit. Shown below is GFP expression in HT1080 cells transduced with the Lenti-miR Virus Library.

Kit Components

Online Resources

 

Technical References

microRNA phenotypic screens

Voorhoeve PM, le Sage C, Schrier M, Gillis AJ, Stoop H, Nagel R, Liu YP, van Duijse J, Drost J, Griekspoor A, Zlotorynski E, Yabuta N, De Vita G, Nojima H, Looijenga LH, Agami R. A genetic screen implicates miRNA-372 and miRNA-373 as oncogenes in testicular germ cell tumors. Cell 2006 Mar 24;124(6):1169-81.

Huang Q, Gumireddy K, Schrier M, le Sage C, Nagel R, Nair S, Egan DA, Li A, Huang G, Klein-Szanto AJ, Gimotty PA, Katsaros D, Coukos G, Zhang L, Puré E, Agami R.
The microRNAs miR-373 and miR-520c promote tumour invasion and metastasis.
Nat Cell Biol. 2008 Feb;10(2):202-10.

Vector-based microRNA expression

Mathijs Voorhoeve, P., C.L. Sage, M. Schrier, A.J.M.Gillis, H. Stoop, R.Nagel, Y. Liu, J.V.Duijse, J. Drost, A. Griekspoor, E. Zlotorynski, N. Yabuta, G. D.Vita, H. Nojima, L.H.J.Looijenga, and R. Agami (2006). “A Genetic Screen Implicates miRNA-372 and miRNA-373 As Oncogenes in Testicular Germ Cell Tumors.” Cell, 124, 1169-1181

Shin, K. J., E. A. Wall, J. R. Zavzavadjian, L. A. Santat, J. Liu, J. I. Hwang, R. Rebres, T. Roach, W. Seaman, M. I. Simon and I. D. Fraser (2006). "A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression." PNAS U S A, 103(37): 13759-64.

Stegmeier, F., G. Hu, R. J. Rickles, G. J. Hannon and S. J. Elledge (2005). "A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells." PNAS U S A, 102(37): 13212-7.

Primary microRNA processing

Han J, Lee Y, Yeom KH, Kim YK, Jin H, Kim VN. “The Drosha-DGCR8 complex in primary micorRNA processing”. Gene Develop, 18: 3016-3027, 2004.

Han J, Lee Y, Yeom KH, Nam JW, Heo I, Rhee JK, Sohn SY, Cho Y, Zhang BT, Kim VN. “Molecular basis for the recognision of primary microRNAs by the Drosha-DGCR8 complex”. Cell, 125: 887-901.2006.

Zeng Y, Yi R, Cullen BR. “Recognition and cleavage of primary microRNA precursors by the nuclear processing enzyme Drosha”. EMBO J, 24:138-148, 2005.

Zeng Y, Cullen BR. “Efficient processing of primary microRNA hairpins by Drosha requires flanking nonstructured RNA sequences”. J Bio Chem, 280:27595-27603, 2005.


microRNA functional study

Bennasser, Y., S. Y. Le, M. L. Yeung and K. T. Jeang (2004). "HIV-1 encoded candidate micro-RNAs and their cellular targets." Retrovirology, 1(1): 43.

Esau C, Kang X, Peralta E, Hanson E, Marcusson EG, Ravichandran LV, Sun Y, Koo S, Perera RJ, Jain R, Dean NM, Freier SM, Bennett CF, Lollo B, Griffey R. MicroRNA-143 regulates adipocyte differentiation.
J Biol Chem. 2004 Dec 10;279(50):52361-5.

Hariharan, M., V. Scaria, B. Pillai and S. K. Brahmachari (2005). "Targets for human encoded microRNAs in HIV genes." Biochem Biophys Res Commun, 337(4): 1214-8.

Johnson, S. M., H. Grosshans, J. Shingara, M. Byrom, R. Jarvis, A. Cheng, E. Labourier, K. L. Reinert, D. Brown and F. J. Slack (2005). "RAS is regulated by the let-7 microRNA family." Cell, 120(5): 635-47.

Kim, V. N. (2005). "Small RNAs: classification, biogenesis, and function." Mol Cells, 19(1): 1-15.

Lee, R. C., R. L. Feinbaum and V. Ambros (1993). "The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14." Cell, 75(5): 843-54.

Lee, Y., K. Jeon, J. T. Lee, S. Kim and V. N. Kim (2002). "MicroRNA maturation: stepwise processing and subcellular localization." Embo J, 21(17): 4663-70.

Olsen, P. H. and V. Ambros (1999). "The lin-4 regulatory RNA controls developmental timing in Caenorhabditis elegans by blocking LIN-14 protein synthesis after the initiation of translation." Dev Biol, 216(2): 671-80.

Pfeffer, S., M. Zavolan, F. A. Grasser, M. Chien, J. J. Russo, J. Ju, B. John, A. J. Enright, D. Marks, C. Sander and T. Tuschl (2004). "Identification of virus-encoded microRNAs." Science, 304(5671): 734-6.

Yi, R., Y. Qin, I. G. Macara and B. R. Cullen (2003). "Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs." Genes Dev, 17(24): 3011-6.

microRNA discovery

Lee RC, Feinbaum RL, Ambros V. “The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14”. Cell, 75:843-854, 1993.

Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvits HR, Ruvkun G. “The 21-nucleotides let-7 RNA regulates developmental timing in Caenorhabditis elegans”. Nature, 403:901-906, 2000.

Lagos-Quintana M, Rauhut R, Lendeckel W, Tuschl T. “Identification of novel genes coding for small expressed RNAs”. Science, 294:853-858, 2001.
 
microRNA targets prediction

Rhoades MW, Reinhart BJ, Lim LP, Burge CB, Bartel B, Bartel DP. “Prediction of plant microRNA targets”. Cell, 110:513-520, 2002.

Lewis BP, Burge CB, Bartel DP. “Conserved seed pairing, often flanking by adenosines, indicates that thousands of human genes are microRNA targets”. Cell, 120:15-20, 2005.

John, B., C. Sander and D. S. Marks (2006). "Prediction of human microRNA targets." Methods Mol Biol, 342: 101-13.

Pooled Lenti-miR virus library Quantity Product Size Catalog # Price MicroRNA Precursor Pooled Library (Pre-packaged) >10^7  IFUs/aliquot 4 virus aliquots PMIRHPLVA-1 $3,850.00 MicroRNA Precursor Pooled Library (HT, Pre-packaged) >10^7  IFUs/aliquot 10 virus aliquots PMIRHPLVAHT-1 $7,800.00

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