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The use of shRNAs holds advantages upon siRNA technology

- It allows transduction of a wide range of cell types, including non-dividing and primary cell lines
- It produces an effective long term gene knock-down effect

The TDI has the capacity to run shRNA screens using both pooled and arrayed libraries. Our expertise are applied to the production and use of high quality ready-to-use viral particles (from glycerol stocks) using several shRNA libraries available on the market but also can support the use and application of other commercially available libraries such as THERMO Decode pooled lentiviral shRNA and SIGMA MISSION® LentiPlex® Pooled shRNA Libraries (viral particles).

The TDI benefits from 2 complete shRNA libraries

1.THERMO Human and Mouse GIPZ Lentiviral shRNAmir Individual Clone libraries

 - Bacterial glycerol stock - utilised to produce pooled or arrayed libraries

2.   Decode pooled lentiviral shRNA whole human genome library

 - Ready to use lentiviral particles

The TDI benefits from an established network off collaborations between academia and industry and is able to support in a collaborative fashion the design of shRNA screenings; the production of lentiviral particles and downstream to the analysis of the NGS data.

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1. Thermo Scientific Human and Mouse GIPZ shRNA Lentiviral Libraries

Human/Mouse GIPZ Lentiviral shRNAmir Individual Clone libraries are used for developing unbiased genome-wide arrayed screens but can also produce customised pooled/arrayed libraries of specific gene families of interest.

Library plates are received in 96 well format as bacterial glycerol stocks. The individual clones are grown, shRNA DNA constructs are isolated and lentiviral particles produced either in a pooled or arrayed fashion.

Genome wide GIPZ Lentiviral shRNAmir Individual Clone libraries allow transient or long-term gene suppression targeting human and mouse genes. Between 3 and 6 shRNA constructs allow gene knockdown of at least 70% (for one out of three constructs) at the mRNA level. TurboGFP tag and puromycin resistance cassette allows positive cell  identification.

GIPZ lentiviral shRNA are provided as individual human or mouse constructs, target gene sets, transfection or transduction starter kits and arrayed libraries. Positive and negative GIPZ shRNA controls are available in glycerol stock and viral particle formats.

1.1 Human GIPZ Lentiviral shRNAmir Individual Clone (Glycerol stock, Cat# RHS6037)

Species: Human 
Library composition: shRNAmir in glycerol stock
Library coverage/representation: 18205 genes/95703 constructs
Promoter hairpin: hCMV
Marker: TurboGFP ex: 482 nm; em: 502 nm
Antibiotic resistance: Puromycin
Lentiviral system: 2nd generation only

Gene Family

# Genes

# Constructs

 Cat #

Human Ubiquitin Conjugation Subset 1

88

599

RHS6040

Human Ubiquitin Conjugation Subset 2

113

812 

RHS6041

Human Ubiquitin Conjugation Subset 3

370 

2419

RHS6042 

Human Phosphatase

254 

1561 

RHS6038 

Human Ion Channel

347 

1884 

RHS6045 

Human GPCR

382 

2591 

RHS6043 

Human Protease

478 

2559 

RHS6044 

Human Protein Kinase

709 

4675 

RHS6039 

Human Drug Targets

4753 

25354 

RHS6046 

Human Rest of Genome

10711 

53249 

RHS6047 

Human Whole Genome

18205 

95703 

RHS6037 

1.2 Mouse GIPZ Lentiviral shRNAmir Individual Clone (Glycerol stock, Cat# RMM5829)

Species: Mouse
Library composition: shRNAmir in glycerol stock
Library representation: 110000+ constructs
Promoter hairpin: CMV
Marker: TurboGFP ex: 482 nm; em: 502 nm
Antibiotic resistance: puromycin
Lentiviral system: 2nd generation only

Link to Technical Manual

TDI HTS Facility ShRNA and cDNA Core Facility

Price Schedule (Effective 17/02/2014)

Service/Item

Price

Format

Individual shRNA clones

£31.87/shRNA construct
(Price on application for large numbers)

Glycerol stock

Non-silencing shRNA control

£31.87

Glycerol stock

pGIPZ Empty Lentiviral vector

£31.87

Glycerol stock

shRNA GAPDH positive control

£31.87/shRNA construct

Glycerol stock

Individual cDNA clones

£31.87/ construct

Glycerol stock

shRNA Request form

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2. Thermo Scientific Decode Pooled Lentiviral shRNA Screening Libraries

Thermo Scientific Decode Pooled Lentiviral shRNA Screening Libraries are pools of GIPZ shRNAs targeting human genes, provided as concentrated lentiviral particles. Through RNAi-mediated silencing of hundreds or thousands of genes in parallel, a pooled lentiviral shRNA screen can be performed to identify genes that regulate cellular responses and signaling pathways, or to discover novel gene functions. In contrast to the costly automated techniques that are required to screen using individually arrayed RNAi reagents, Decode pooled shRNA screening libraries allow the researcher to transduce and screen a population of cells within a few tissue culture dishes.

Decode pooled lentiviral shRNA - Human genome

Species: Human
Library composition: lentiviral particles (10 pools of 9570 shRNA/pool) - 18205  genes
Promoter hairpin: hCMV
Marker: TurboGFP ex: 482 nm; em: 502 nm
Antibiotic resistance: Puromycin

Link to ThermoScientific Decode Pooled Lentiviral shRNA Screening Libraries