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shRNA and ORFeome Core

shRNA and ORFeome Core

The discovery of short hairpin RNAs (shRNAs) suppressing gene expression in mammalian cells enables large-scale loss-of-function screens by using genome-wide shRNA libraries. Genome scale over-expression libraries allow large-scale gain-of-function screens. The ShRNA and ORFeome core makes available to the MDACC research community the human and mouse shRNA libraries, human ORFeome collaboration and LentiORF libraries (http://www.thermoscientificbio.com/openbiosystems/). The core facility can provide individual shRNA plasmids and cDNA clones for MDACC laboratories, or carry out a screening experiment using the libraries. Purchase of clones or libraries through the shRNA and ORFeome core reduces costs and decreases turnaround time for researchers.

Location:

The UT MD Anderson Cancer Center
Department of Molecular and Cellular Oncology
1515 Holcombe Blvd, Room Y8.5719, Houston, TX 77030

Available Libraries:

Human and mouse ShRNA libraries: GIPZ Lentiviral shRNAmir libraries targets the entire human and mouse genome with multiple shRNAmir per target gene.
For details, please visit: http://www.thermoscientificbio.com/rnai-and-custom-rna-synthesis/shrna/gipz-lentiviral-shrna/

Human ORFeome Collaboration: each clone contains a sequence-verified open reading frame (ORF) subcloned into Gateway™ recombinational entry vector.
For details, please visit: http://www.thermoscientificbio.com/gene-expression-cdnas-orfs/mammalian-orfs/orfeome-collaboration-collection

Human Precision LentiORFs: an expression-ready library, lentiviral ORFs.
For details, please visit: http://www.thermoscientificbio.com/mammalian-orf/precision-lentiorfs/?Parent=12884901918

Pre-Assembled Sub-libraries:

  • Human Kinase shRNA sub-libraries (40 96-well plates)
  • Human Phosphatase shRNA sub-libraries (15 96-well plates)
  • Human E3 ligase (BTB) shRNA sub-library (9 96-well plates)
  • Human E3 ligase (Ring) shRNA sub-library (15 96-well plates)
  • Human E3 ligase (DDB1-like U box F box SOCS box HECT ZNF_A20) shRNA sub-library (8 96-well plates)

Services**:

  • Individual Clone distribution
  • Customized shRNA or ORF libraries
  • DNA purification: Facilitated by the Tecan automated workstation, the core will purify DNA in a high throughput manner.
  • Lentiviral production and infection

**The shRNA and ORF reagents are available to MDACC researchers ONLY due to licensing agreements. If you have any questions, please contact Dr. Yutong Sun at ysun2@mdanderson.org.

Personnel

Director:

Mien-Chie Hung, Ph.D.
Vice President for Basic Research
Professor and Chair
Department of Molecular and Cellular Oncology- Unit 108
Professor, Department of Surgical Oncology
Distinguished Teaching Professor
Ruth Legett Jones Distinguished Chair
Email: mhung@mdanderson.org
Tel: 713-792-3668
Fax: 713-794-3270

Co-directors:

Dihua Yu, M.D., Ph.D.
Hubert L. & Olive Stringer Distinguished Chair in Basic Science
Professor and Deputy Chair
Department of Molecular and Cellular Oncology
Co-Director, Center of Biological Pathways
Email: dyu@mdanderson.org
Tel: 713-792-3636
Fax: 713-792-4454

Yutong Sun, Ph.D.
Assistant professor
Email: ysun2@mdanderson.org
Tel: 713-792-3677
Fax: 713-794-3270

Research Scientist:

Longfei Huo, Ph.D.
Email: Lhuo@mdanderson.org
Tel: 713-795-0524
Fax: 713-794-3270

Equipment

The shRNA and ORFeome Core offers a few cutting-edge tools tailored for high-throughput screens. To schedule the equipment usage, please contact the core facility directly at ShRNAandORFeomeCoreFacility@mdanderson.org,  Dr. Yutong Sun at ysun2@mdanderson.org, or Dr Longfei Huo at lhuo@mdanderson.org.

An automated workstation EVO100 (Tecan) is located in a laminar flow hood, which can facilitate high throughput tissue culture.

 

 

 

 

An automated workstation EVO200 (Tecan) is equipped with both span8 and MCA96 liquid handling arms, a vacuum and an orbital shaker, which allows automated liquid handling, plasmid purification and manipulation in a high throughput manner.

 

 

 

Biotek synergy H4 plate reader is capable of performing absorbance, fluorescence and luminescence assays.

 

 

 

 

 

High-Content Imaging Systems:

ImageXpressMicro (Molecular Devices) is a widefield high content microscope capable of providing automated cellular imaging in fluorescent, transmitted light, and phase-contrast modes for fixed- or live-cell assays. It is also equipped with Environmental Control that allows for time-lapse imaging of live cells.

 

 

ImageXpressUltra is an automated point-scanning confocal microscope.
Both ImageXpressMicro and ImageXpressUltra  are able to perform automated acquisition and analysis of imagines in multi-well plates (up to 384-well plates).

 

 

The high content imaging systems enable investigators to perform image-based high throughput screens, measuring critical events including apoptosis, cell cycle, micronuclei, neurite outgrowth, angiogenesis, intracellular translocation, endocytosis, cell signaling, and multi-wavelength cell scoring.


© 2014 The University of Texas MD Anderson Cancer Center