Animal Gene Editing Unit
                       
    Main Page   Staff Directory   Publications   RESOURCES      
                       
   
a filler
  Weiping YU  
  Lab Location: #4-01

email:
yu_wei_ping@brc.a-star.edu.sg
tel: 64070684
 
a a a
a a a
  Key Publications  
 

Wei Zhang, Selvaratnam Thevapriya, Paul J. Kim, Wei-Ping Yu, H. Shawn Je, Eng King Tan and Li Zeng  (2014)
Amyloid precursor protein regulates neurogenesis by antagonizing miR-574-5p in the developing cerebral cortex.
Nature Communications 5:3330

Yuen-Peng Tan, Shaobing Li, Xiao-Juan Jiang, Wailin Loh, Yik Khon Foo, Chay-Boon Loh, Qiurong Xu, Wai-Hong Yuen, Michael Jones, Jianlin Fu, Byrappa Venkatesh, Wei-Ping Yu  (2010)
Regulation of protocadherin gene expression by multiple neuron-Restrictive silencer elements scattered in the gene cluster.
Nucleic Acids Research 38:6985-4997

Xiao-Juan Jiang, Shaobing Li, Byrappa Venkatesh, Wei-Ping Yu (2009)
Identification and comparative analysis of the protocadherin cluster in a reptile, the green anole lizard.
PLoS ONE  4:e7614 

Wei-Ping Yu*, Viknesawari Rajasegaran, Kenneth Yew, Wai-lin Loh, Boon-Hui Tay, Chris T. Amemiya, Sydney Brenner*, Byrappa Venkatesh*  (2008)
Elephant shark sequence reveals unique insights into the evolutionary history of vertebrate genes: A comparative analysis of the protocadherin cluster.
Proc Natl Acad Sci U S A 105:3819-3824 (*Correspondence)

Wei-Ping Yu*, Kenneth Yew, Vikneswari Rajasegaran, Byrappa Venkatesh*.  (2007)
Sequencing and comparative analysis of fugu protocadherin clusters reveal diversity of protocadherin genes among teleosts.
BMC Evolutionary Biology 7:49 (*Correspondence)

Wei-Ping Yu, Sydney Brenner and Byrappa Venkatesh.  (2003)
Duplication, degeneration and complementation of the nested synapsin-Timp genes in Fugu rubripes.
Trends in Genetics 19:180-183

Wei-Ping Yu, Catherine J. Pallen, Alice Tay, Frank R. Jirik, Sydney Brenner, Y.H. Tan, and Byrappa Venkatesh.  (2001)
Conserved synteny between the Fugu and human PTEN locus and the evolutionary conservation of vertebrate PTEN function.
Oncogene 20:5554-5561

Wei-Ping Yu, Ellen J. Collarini, Nigel P. Pringle, and William D. Richardson.  (1994)
Embryonic expression of myelin genes: evidence for a focal source of oligodendrocyte precursors in the ventricular zone of the neural tube.
Neuron  12:1353-1362

 

 

 

 
a a a
 
  Weiping YU


Weiping Yu received his BSc from Nankai University in 1983, MSc from Peking Union Medical College in 1986, and his Ph.D in Neurobiology from University College London in 1995. He then moved to IMCB for postdoctoral training first with Prof Catherine Pallen and later with Prof Byrappa Venkatesh. In 2003, he set up his own lab at the National Neuroscience Institute (NNI) where his research focuses on study of gene regulation mechanisms involved in the neurodegenerative diseases. In 2011, He moved to Biological Resource Centre (BRC), A*STAR, helping set up a service oriented Animal Gene Editing Laboratory (AGEL). In 2015, he took a joint position (while his primary employment is with BRC) and re-joined IMCB as a Group Leader in the Infrastructure, Technology & Translational Division.

     
  Animal Gene Editing Laboratory (AGEL)
 


Genetically modified animals are powerful tools for studying physiological function of genes in vivo in the whole animal context. The primary goal of AGEL is to provide services in creating genetically modified animal models and to conduct research on developing new gene editing technologies and novel animal models for human diseases. AGEL provides a wide spectrum of gene editing services in animals, including creation of transgenic rodents by pronuclear injection and gene knockin/knockout in mice via ES cell targeting. AGEL employs a number of gene editing methods including the recently developed TALEN and CRISPR/Cas9 nuclease-based techniques. We provide a comprehensive range of services that cover all of the steps involved in generating genetically modified animals including design of gene targeting strategy, synthesis and preparation of reagents and production of F1 heterozygotes etc.

Services Provided by AGEL

1) Gene targeting via ES cells

    • Design and construction of gene targeting vector
    • Generation of gene targeted ES cell lines
    • Creation of knockout/knockin chimeras by blastocyst injection
    • Production of F1 heterozygotes (germline transmission)

For floxed conditional knockout mice, we strongly advise researchers to search the IMPC database (https://www.mousephenotype.org/) to find out if the consortium has already generated targeted ES cell lines or knockout animals for the gene of interest.  Researchers may directly order targeted ES cells from the consortium, and AGEL can generate the knockout animals by blastocyst injection.  

2) Rodent transgenesis

    • Construction of tissue specific promoter driving transgene expression cassette
    • BAC engineering:  insertion of reporter gene, exon deletion or single/multiple nucleotide mutations
    • Stable transfection and screening for ES cells containing the genetically modified BAC
    • Creation of transgenic animals by pronuclear injection or blastocyst injection (BAC transgenesis)

Pronuclear injection of transgene construct is the traditional way to overexpress a protein coding gene or to express a reporter gene under a tissue-specific promoter in animals. However, the expression pattern of the transgene in animals generated by this method can be highly variable from line to line, and from generation to generation, due to random integration (positional and copy number) of the transgene. These issues occur less frequently (but are not completely eliminated) with BAC transgenesis.

3) Transgene expression at Rosa26 locus

    • Cre-mediated tissue/lineage-specific transgene expression at Rosa26
    • Tetracycline inducible transgene (protein-coding gene or shRNA) expression at Rosa26 with Cre-mediated tissue/lineage specificity

As a more precise alternative to pronuclear injection of expression constructs, we have developed a transgene expression platform that allows for a transgene cassette to be integrated as a single copy specifically at the Rosa26 locus. The expression of the transgene is under the control of a ubiquitous CAGGS promoter with a preceding floxed transcriptional stop signal inserted between the promoter and the transgene coding sequence. Tissue/lineage specificity of transgene expression is achieved by Cre-mediated deletion of the floxed “STOP”. Thus, by crossing with lines expressing Cre under the control of a range of tissue-specific promoters one can achieve different patterns of transgene expression with same animal model, ─ one model for multiple studies. Transgene expression can also be controlled temporally by tetracycline induction. The highly controllable (spatial and temporal) and predictable (single copy at predetermined genomic locus) nature of this system makes it ideal for studying the pathogenesis of gene mutations that may cause predisposition to human disease, because individual transgenic lines with different mutations can be directly compared for phenotypic differences. In addition to protein-coding genes, this system also supports shRNA expression under a well characterized miRNA expression cassette.  This facilitates acute knockdown of gene expression at any developmental stage (tetracycline-inducible) and in any tissue (with a specific Cre line of choice). Integration of individual transgenes at the docking site in ES cells is mediated by a highly efficient recombinase-mediated cassette change (RMCE) process, which allows for rapid generation of multiple transgenic lines simultaneously.

     

(Click to view larger images)

 

 


4) TALEN & CRISPR/Cas9 mediated gene editing in mice and rats

    • Individual or large scale TALEN plasmid assembly and activity assay
    • TALEN or CRISPR/Cas9 mediated reading frame shift (indel) or single/multiple nucleotide mutagenesis in mice and rats

AGEL provides full support for the creation of genetically modified rodents using TALEN or CRISPR/Cas9 technology, including assembly and construction of specific TALEN plasmids, activity testing of TALENs and gRNA (see RESOURCES), reading frame shift mutagenesis in cells and animals (mice and rats), oligonucleotide-mediated single/multiple nucleotide mutagenesis in mice and rats etc. AGEL’s services are comprehensive, from design of the targeting strategy, synthesis and preparation of the reagents, microinjection of oocytes, to analysis of chimeric founders by deep sequencing and breeding of mutation segregated F1 heterozygotes.


(Click to view larger images)

 

 


5) Ad hoc services: In addition to the services listed above, AGEL provides advice and technical supports for custom’s projects relating to genetically modified animals and all enquiries are welcome. Please contact us AGEL-BRC@brc.a-star.edu.sg and we will get back to you as quickly as possible.