Functional Genomics & RNAi

ZFN Webinar Series

Zinc Finger Nuclease Webinar Series

Everything you wanted to know about genome editing but didn’t know who to ask!

Join Sigma Life Science® for a series of free, live online seminars based upon the zinc finger nuclease technology. Each seminar will be presented by an expert on the topic, followed by a question and answer period. Attend the seminar from the comfort of your own desk.

If you cannot attend a seminar at the provided date, don’t worry. The online seminars will be recorded and hosted live on this website. So, if you have to miss the live seminar, or need to review the information, you can view the seminar at any time that is convenient for you.

 

Previous Webinars:

May 6, 2010 Introduction to Zinc Finger Nuclease Technology
Presented by Trevor Collingwood, Ph.D.

Zinc finger nucleases (ZFNs) are a class of engineered DNA-binding proteins that enable manipulation of the genome with unprecedented ease and precision. ZFNs create targeted double-strand breaks (DSBs) in the genome at user-specified locations. These DSBs are repaired through the cell's natural DNA-repair processes, namely homologous recombination and non-homologous end joining. These DSB-caused endogenous processes are harnessed to generate precisely targeted genomic edits resulting in both cell lines and organisms with targeted gene deletions integrations, or modifications. In this webinar, we will discuss the design, mechanism of action, and various applications of ZFN technology.



May 27, 2010 Zinc Finger Nuclease Applications: Gene Knockout in Cell Lines
Presented by Trevor Collingwood, Ph.D.

Rational genome engineering in mammalian cells is of enormous potential across basic research, drug-discovery as well as cell-based assays. Zinc finger nucleases (ZFNs) are a class of engineered DNA-binding proteins that facilitate targeted editing of the genome by creating double-strand breaks (DSBs) at user-specified locations. Within these chimeric proteins the DNA binding specificity of the zinc finger protein determines the site of nuclease action. Such engineered ZFNs are able to recognize and bind to a specified locus and evoke a DSB in the targeted DNA with high efficiency and precision. ZFN-induced double-strand breaks are typically repaired by an imperfect cellular repair mechanism called non-homologous end joining (NHEJ). Due to the low fidelity of NHEJ, a subset of DSBs within a ZFN-treated cellular population will be misrepaired, leading to variable genetic insertions or deletions at the site of the DSB. Drawing from our work with transformed cell lines, primary human cells, and multi-potent stem cells, we will present examples of both single and multiple gene knockouts.



June 15, 2010 Zinc Finger Nuclease Applications: Targeted Gene Integration
Presented by Trevor Collingwood, Ph.D.

Zinc finger nucleases (ZFNs) are a class of engineered DNA binding proteins that facilitate efficient targeted editing of the genome by creating double-strand breaks (DSBs) at user-specified locations. The cell then employs the natural DNA repair processes of either homology-directed repair (HDR) or non-homologous end joining to heal the targeted break. HDR enables insertion of a transgene or other defined alterations into the targeted region. By this approach, a donor template is used that contains the transgene flanked by sequences that are homologous to the regions either side of the cleavage site. This donor is co-delivered into the cell along with the ZFNs. Target integration can be performed at user-specified genomic locations - including transgene expression from safe harbor loci. Utilization of a safe harbor locus for targeted integration enables stable expression of the transgene without adverse effects arising from the site of integration with decreased experimental variability. ZFNs are also used for gene tagging whereby a marker, such as GFP, can be fused directly to the endogenous gene. These types of experiments are used for the study of gene expression and cellular localization of the protein. Additionally, ZFNs can be used to recreate or correct disease–related mutations at the endogenous locus. In this webinar, we will discuss the various applications of gene integration.



Advanced topics to follow, Autumn 2010

Trevor Collingwood, Ph.D.

Dr. Collingwood is Manager of Technology Research at Sigma Life Sciences. He gained his PhD in biochemistry at the University of Waikato, New Zealand, followed by post-doctoral studies on gene regulation at Cambridge University, UK. He then moved to the US National Institutes of Health in Bethesda as a visiting scientist where he continued his work on the role of chromatin structure in the control of gene expression. In 2000, Dr Collingwood joined Sangamo BioSciences in California where he led the Enabling Technologies group, focusing on development of the zinc finger nuclease platform for genome engineering. In 2008 he moved to his current position at Sigma where his work continues to focus on technologies for the control of gene expression.

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