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Home |  RNAi | miR-express Lentiviral mic (2)
microRNA biogenesis
A primary miRNA transcript is encoded in the genome and transcribed as hairpins in the nucleus.  These hairpins are initially processed by the enzyme Drosha/Pasha (DGCR8) and exported into the cytoplasm where it is further processed by Dicer into mature miRNA duplexes. After strand separation, the mature miRNA represses protein synthesis either by blocking translation or causing transcript degradation.
Purchasing Information
Cat # Description Literature ACA COM Buy
HMR4842
 miR-express human lentiviral microRNA individual clone - glycerol stock   $450 $450 Click to show search field
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(only clones from HMR4842)
HMR4843 pLemiR lentiviral empty vector-glycerol stock   $300 $300
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HMR4867
 miR-express Non-silencing Control - glycerol stock   $300 $300
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HMR5152 miR-express human lentiviral microRNA library-Glycerol   call call
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miR-express Lentiviral microRNA
for over-expression of human pri-miRNA transcripts
The miR-express™ lentiviral microRNA (miRNA) library is a collection of individual miRNA precursors cloned into a lentiviral vector allowing efficient over-expression of human primary microRNA (pri-miRNA). This collection was created in collaboration with Dr. Muneesh Tewari at the Fred Hutchinson Cancer Research Center. The goal of this project was to clone all known human pri-miRNA transcripts with their natural flanking sequences in lentiviral vectors.

Human miRNAs were selected from the Sanger miRbase database version 11.0 (http://microrna.sanger.ac.uk/sequences) and the genome coordinates were used as a reference to design primers to PCR amplify the pri-miRNA sequences with 5’ and 3’ flank sequences.  The primers for amplification of all human primary miRNAs were designed to retain as much flanking sequence as possible for efficient Drosha processing and endogenous regulation. On average 200 bases of flanking sequence was retained on both sides of the precursor miRNA. The pri-miRNA inserts were then cloned into the lentiviral vector pLemiR™, under the control of a CMV promoter.
 
Highlights of the miR-express collection include:
  • miRNA transcripts are in their native sequence context to ensure endogenous RNAi processing into mature miRNAs
  • miRNAs are expressed in target cells through transfection or transduction
  • TurboRFP allows tracking of miRNA expression
  • Puromycin selectable marker
  • Lentivirus-based system for expression in a wide range of cell lines including primary and non-dividing cells
  • Transient, stable and in vivo studies are possible
Additional technical details

Useful links:
miRbase at the Sanger Institute

Efficient processing of pri-miRNA precursors into mature miRNAs
miR-express pri-miRNA-302a are processed into mature miRNA duplexes in HEK293 cells which dont normally express miR-302a

Please note that the pLemiR vector is not compatible with third generation packaging systems such as ViraPower™ from Invitrogen. We recommend the Trans-Lentiviral™ Packaging System for use with our vectors.

Finding your microRNA of interest

  1. Enter microRNA ID, accession, symbol(s), or catalog numbers into the gene search. Results will be returned in a tabbed format. Click on shRNA under the shRNAs/RNAi tab.
  2. Click on the oligo ID link for additional cloning and sequence information. Complete insert sequence is displayed. Precursor miRNA sequence can be obtained from the datafile in the purchase grid or from miRbase.
  3. Order directly online
Shipping Information:

Individual miR-express microRNA constructs are shipped as glycerol stock cultures on wet ice. Open Biosystems checks all cultures for growth prior to shipment. Store at -80ºC.

References:
miRBase: microRNA sequences, targets and gene nomenclature.
Griffiths-Jones S, Grocock RJ, van Dongen S, Bateman A, Enright AJ.
NAR, 2006, 34, Database Issue, D140-D144
The microRNA Registry.
Griffiths-Jones S.
NAR, 2004, 32, Database Issue, D109-D111
A uniform system for microRNA annotation.
Ambros V, Bartel B, Bartel DP, Burge CB, Carrington JC, Chen X, Dreyfuss G, Eddy SR, Griffiths-Jones S, Marshall M, Matzke M, Ruvkun G, Tuschl T.
RNA, 2003, 9(3), 277-279
Stegmeier F et al (2005) "A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells" PNAS Vol 102:37; 13212-13217
Shimada, Takashi, et. al,  Development of Vectors Utilized for Gene Therapy for AIDS, AIDS 4, 1995

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