shMIMIC Lentiviral microRNA Pooled Libraries

Perform functional screening of hundreds or thousands of microRNAs without high-throughput automation


shMIMIC Lentiviral microRNA Pooled Libraries
A pooled lentiviral microRNA screen can be performed to identify microRNAs that regulate development and signaling pathways, without the need for costly high-throughput automation.

Optimal microRNA expression for high-confidence functional analysis screening results

A novel construct for stable expression of a mature microRNA, shMIMIC Lentiviral microRNA pooled libraries are available to target every human and mouse microRNA described in the miRBase database (www.mirbase.org).

Pooled lentiviral libraries offer an efficient and cost-effective method for screening large numbers of microRNAs without automation. shMIMIC pooled libraries are offered for all human and mouse miRBase matures, as well as a Human/Mouse Conserved collection.

Library construction, pooling techniques, and high-throughput sequencing-compatible screening workflows have been experimentally validated to ensure reproducibility and accurate hit identification. Learn about optimal library design and the pooled screening workflow on our Pooled Lentiviral Screening Libraries page.

Highlights:

You may also select shMIMIC Inducible microRNA pooled libraries for tight regulation of expression from the Tet-On® 3G transactivator protein

Each shMIMIC lentiviral library includes:

  • 30 Non-targeting controls
  • 80 species-specific positive control shRNAs targeting 10 common viability and reference genes (8 shRNAs each)
  • A data file including construct sequences, miRBase gene IDs, and counts per millions of mapped reads
1Total constructs includes 110 non-targeting and gene-specific shRNA constructs. All shMIMIC Lentiviral microRNA pooled libraries are ≥ 5 × 108 TU/mL (+/- 20 %) provided in 8 tubes × 25 µL (200 µL total)
shMIMIC microRNA Pooled Library Number of mature microRNAs Number of unique designs Number of pools × number of constructs per pool1
Human 2580 2555 1 pool × 2665 constructs
Mouse 1913 1896 1 pool × 2006 constructs
Human-Mouse Conserved 355 human + 341 mouse 386 1 pool x 496 constructs

Custom collections of microRNA pooled libraries are available

  • Request a custom collection of human, mouse, or rat microRNAs
  • Available pool sizes are between 50 to 10,000 constructs
  • Pools are provided as purified, concentrated lentiviral particles
  • ≥ 5 x 108 TU/mL for constitutive vectors and ≥ 1 x 107 TU/mL for inducible vectors

Request pricing for a custom pooled library

Products recommended in our validated protocol:

Before ordering, download these valuable tools to carefully plan your pooled lentiviral screen and calculate the amounts of components required:

Important Notice

The shMIMIC Lentiviral microRNA Pooled Libraries are solely for internal research use (as set forth in the Product Terms and Conditions) in laboratories where the containment measures stated below and in applicable laws and regulations are met. Products may not be used for diagnostic, therapeutic or other commercial purposes and may not to be administered to humans for any purpose or to animals for therapeutic purposes. SMARTvector Lentiviral shRNA Pooled Libraries are provided as lentiviral particles are replication-incompetent, self-inactivating (SIN) and non-pathogenic (do not cause infectious human disease).

Any investigator who purchases lentiviral particle products is responsible for consulting with their institution's health and biosafety personnel for specific guidelines on the handling of lentiviral vector particles. Furthermore, each investigator is fully responsible for obtaining the required permissions for research using and the acceptance of replication-incompetent SIN lentiviral vectors and replication-defective lentiviral particles into their local jurisdiction and institution.

  
HazardousNo
Shelf Life12 Months
Shipping ConditionDry Ice
Storage Condition-80 C
Figure 1.jpg

Elements of the shMIMIC microRNA Lentiviral Backbone

Figure 1.jpg

Vector Element Utility
5' LTR 5' Long Terminal Repeat necessary for lentiviral particle production and integration of the construct into the host cell genome
Ψ Psi packaging sequence allows viral genome packaging using lentiviral packaging systems
RRE Rev Response Element enhances titer by increasing packaging efficiency of full-length viral genomes
tGFP or tRFP TurboGFP or TurboRFP reporter for visual tracking expression upon doxycycline induction
None No-reporter option for use in applications where fluorescence is not required or desired
IRES Internal Ribosomal Entry Site allows expression of fluorescent marker and puromycin resistance in a single transcript
PuroR Puromycin resistance permits antibioticselective pressure and propagation of stable integrants
SMARTvector universal scaffold Optimized proprietary scaffold in which mature microRNA sequence is embedded
WPRE Woodchuck Hepatitis Post-transcriptional Regulatory Element enhances transgene expression in target cells
3' SIN LTR 3' Self-inactivating Long Terminal Repeat for increased Lentiviral safety


Pooled-shRNA-screening-workflow

shMIMIC Pooled Screening Workflow

Pooled-shRNA-screening-workflow

Assay Development and Optimization: Establish optimal experimental conditions, including those for a) lentiviral transduction and b) screening parameters, such as selective pressure and time between collection of reference and experimental samples.
Primary Screen: A stable population of cells expressing single integrands of constructs are created by transducing lentiviral pools at low MOIs. Transduced cells are then split into reference and experimental populations for application of a selective pressure that induces the phenotype of interest. Genomic DNA (gDNA) is then isolated from reference and experimental populations of transduced cells. Illumina-adapted primers and Phusion Hot-Start II High Fidelity DNA Polymerase are used to PCR amplify integrated construct sequences and add Illumina flow-cell binding sequences. The resulting amplicons are run on Illumina platform sequencers, using the sequencing primers provided.
Hit Identification and Follow-up: Construct sequences are identified in reference and experimental libraries. Constructs that are enriched or depleted during the screen are identified as hits, and the genes that they target are identified. Hits can be confirmed and studied further using individual constructs that can be ordered from the Dharmacon catalog collection.


variable-promoter-strength-activity

Strength of promoter activity varies from one biological context to another and can affect experimental outcomes.

variable-promoter-strength-activity

Promoter activity varies across several human and rodent cell lines. Cells were plated at a density of 50,000 cells per well in a 24-well plate and transduced at MOI = 15 with SMARTvector Empty Vector Control Particles expressing TurboGFP. Promoter activity was assessed at 72 hours post-transduction by the fluorescence intensity of TurboGFP.


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The Human-Mouse Conserved microRNA Collection

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The Human-Mouse Conserved microRNA pooled library is useful for researchers that need ultra-high fold coverage for subtle phenotypes, or who may not have enough cells (e.g. primary cells) for a larger screen and wish to focus on highly conserved microRNAs. It can be used in either human or mouse models, and translates well to in vivo applications.
It includes 386 total mature microRNAs:
302 human and mouse matures that are identical
35 mouse matures in broadly conserved families that are not identical to human
49 human matures in broadly conserved families that are not identical to mouse
Broadly conserved families as defined at www.targetscan.org