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   Today: Last Update: 01 April 2013


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Arrayed RNAi screens
Boutique Libraries
GIPZ lentiviral shRNAmir Hs - genome-wide, tGFP, non-inducible
GIPZ lentiviral shRNAmir Mm - genome-wide, tGFP, non-inducible
TRIPZ lentiviral shRNAmir - individual shRNAmir, tRFP, inducible
LEX lentiviral protein expression - individual shRNAmir, under development
Pooled screens - under development
FANTOM and RIKEN plasmid libraries

In-house results

LISA maintains human and murine versions of proteincoding focused genome-wide pGIPZ-shRNAmir libraries in a lentiviral vector. Each library comprises some 60,000 individual plasmids which are randomly arrayed across 96 well microplates.

The pGIPZ plasmid encodes a single transcript comprising a puromycin resistance marker, turboGFP, and the shRNAmir. These elements can be packed into lentivirus particles for transduction or the pGIPZ plasmid can be used on its own in transient transfections.

IMB-internal researchers can obtain any of these plasmids as E.coli glycerol stocks or as ready-to-use lentivirus preparations. We are happy to provide just one plasmid or lentiviral supernatant in a tube or entire sets arrayed in 96 well microplates.

For researchers who need an inducible vector or who cannot tolerate the co-expression of GFP LISA offers the empty version of a pTRIPZ-shRNAmir for subcloning. pGIPZ and pTRIPZ possess identical multiple cloning sites and any shRNAmir of interest can be easily transferred by restriction digest. The pTRIPZ plasmid encodes a doxycycline inducible transcript comprising a puromycin resistance marker, turboRFP, and the shRNAmir. <top>

LISA offers bulk lentivirus preparation for any pTRIPZ-shRNAmir plasmid you have created. Currently LISA is unable to offer a cloning service.

For researchers who would like to use lentivirus particles for protein expression or who would like to re-introduce a RNAi resistant protein in their validation studies, LISA has acquired the empty pLEX-MCS lentiviral expression vector.
LEX, GIPZ, and TRIPZ systems all use the same lentiviral packaging system. LISA currently offers pLEX-MCS as DNA for you to subclone your protein of interest. Once returned, LISA can produce bulk lentivirus as with GIPZ and TRIPZ plasmids.

The pLEX-MCS manual can be found here.

Protein expression clones may be found in our FANTOM and RIKEN clone libraries. See here for more.

For researchers who would like to perform RNAi screens, LISA offers small and ready-to-use 'Boutique Libraries' which have been re-arrayed from the original genome-wide pGIPZ-shRNAmir libraries. You can pick from the Boutique Libraries that are already available or you can design your own by specifying the human and/or murine target genes. Read more ...

Researchers who contemplate screening using a pooled instead of an arrayed RNAi library might be interested to know that LISA is currently evaluating the acquisition of a pooled library. <top>

The FANTOM3 set contains 103,000 fully sequenced murine cDNA clones with additional annotation data including Cap-analysis gene expression (CAGE) data, which allows high-throughout gene expression analysis and the profiling of transcriptional start sites, including promotor usage analysis. Gene Identification Signature (CIG) and Genome Signature Cloning (GSC) data is also included which allows identification of mRNA variants and rare transcripts.

The FANTOM2 set consists of the original 21,000 murine FANTOM (Functional Annotation of the Mammalian Genome) clones plus a further 39,000 more selected from the full length cDNAs produced by the Genome Exploration Research Group at RIKEN's Genomic Resource Centre.

The 12,000 RIKEN murine cDNA clones were established by the Genome Exploration Research Group at RIKEN's Genomic Resource Centre using full-length cDNA technology.

The IMB is one of three Australian participants in the FANTOM network next to the Eskitis Institute (Griffith University) and the Flinders Medical Centre. More information available at http://fantom.gsc.riken.jp <top>

FANTOM2 <http://fantom.gsc.riken.jp/2/>
FANTOM3 <http://fantom.gsc.riken.jp/3/>

The clone order form is here.



Examples (Results obtained in-house)