Biomedicum Functional Genomics Unit Libraries

FuGU/Libraries maintains and distributes human and mouse genome-wide, arrayed lentiviral vector -based shRNA (The RNAi Consortium (TRC) library, Broad Institute) and CRISPR/Cas9 gRNA (Sigma-Aldrich) libraries. The shRNA libraries consist of 160,000 distinct constructs, whereas the gRNA libraries contain altogether ~ 80,000 clones. Non-targeting control constructs for both gRNA and shRNA constructs are also available. Main services include the production of glycerol stocks, DNA preparations, standard and concentrated recombinant viral particles from the libraries and customer-provided constructs. Additionally, shRNA/ CRISPR library knockdown/-out validation is done as a service by using LightCycler 480 instrumentation and Universal ProbeLibrary (UPL) probes (Roche). FuGU/Libraries is a non-profit core facility. Reagent, equipment and labor expenses are charged from UH customers, whereas for external customers overheads are charged. Pricing has been done according to the University of Helsinki finances instructions.

Contact person: Principal investigator and genome editing specialist Topi Tervonen, +358 2941 25494, topi.tervonen@helsinki.fi; Research coordinator and virus production specialist Katariina Karjalainen (Maria Salmela on maternity leave), +358 2941 25494, katariina.karjalainen@helsinki.fi

Location: Biomedicum Helsinki 1, Haartmaninkatu 8, Meilahti campus

Services

  • Bacterial glycerol stock preparations from the library constructs
  • Plasmid DNA preparations from the library constructs
  • Standard or concentrated lentiviral particles from the library constructs
  • Standard or concentrated (only lenti) retro- and lentiviral particles from customer constructs
  • qRT-PCR validation, for example, of efficiency of shRNA-mediated knockdown with LightCycler 480 instrument

Resources

  • Arrayed genome-wide lentiviral human shRNA library, 16,000 genes, 5 shRNA constructs/gene
  • Arrayed genome-wide lentiviral mouse shRNA library, 15,950 genes, 5 shRNA constructs/gene
  • Arrayed genome-wide lentiviral human CRISPR/Cas9 gRNA library, 17,166 genes, 2 gRNA constructs/gene
  • Arrayed genome-wide lentiviral mouse CRISPR/Cas9 gRNA library,  20,430 genes, 2 gRNA constructs/gene
  • Universal ProbeLibrary (UPL) from Roche

Instruments

  • Roche LightCycler 480 II
  • SANYO MDF-U73V and Panasonic MDF-C8V1-PE deep freezers (-80ºC) for long-term storage of the libraries

Applications

  • Stable shRNA-mediated gene knockdown by lentiviral gene transfer
  • Stable gene knockout via CRISPR/Cas9 gRNA by lentiviral gene transfer (KO-ready)
  • Efficient gene transfer to hard-to-transfect cells by recombinant lentiviral particle delivery
  • Transient knockdown or permanent knockout in easy-to-transfect cells by transfection of the viral transfer vector (gRNA vectors require Cas9 enzyme from external sources)
  • Generation of user defined libraries
  • Multiplex gene knockout (for example, redundant gene families)

User fees

The research infrastructure is supported by HiLIFE and Faculty of Medicine of University of Helsinki and Biocenter Finland. The academic prices (excl. VAT) can be found from the table below. Please ask for a quotation for you project through this portal.

Product Description Starting price (eur) 1st order

Starting Price (eur) additional orders

Concentrated lentivirus, 180 ul

shRNA from TRC1 library  540 285
  gRNA from CRISPR library    
  Customer plasmid 375 205
Concentrated lentivirus, 380 ul shRNA from TRC1 library 595 340
  gRNA from CRISPR library    
  Customer plasmid 430 260

Concentrated lentivirus sucrose purification

  35 10
Lentivirus MINI TRC1 (gene set x5) 390 205

 

Customer plasmid

205 70

Lentivirus MIDI

shRNA from TRC1 library 315 125
  gRNA from CRISPR library 440 150
  Customer plasmid 225 85
Retrovirus MINI From customer's glycerol stock 165 35
  From customer's plasmid 95 20
Retrovirus MIDI From customer's glycerol stock 195 60
  From customer's plasmid 110 25
Glycerol stock   15 10
DNA preparation MINI 5 x shRNA from TRC1 library 160 115
  2 x gRNA from CRISPR library 95 45
  Customer glycerol stock 65 15
DNA preparation MAXI shRNA from TRC1 library 160 80
  Customer glycerol stock 155 75
Bacterial transformation   55 20
Cell ampouls HeLa/Phoenix Ampho/Phoenix Eco/ HEK-293FT 10