Targeted Genome Editing using Zinc Finger Nucleases Workshop

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	Overview Dates and Locations Lecture and Lab Topics	Previous Guest Lecturer Feedback from Attendees

Overview

Zinc finger nucleases (ZFNs) enable manipulation of the genome with unprecedented ease and precision. ZFNs are a class of engineered DNA-binding proteins that facilitate targeted editing of the genome by creating double-strand breaks (DSBs) in DNA at user-specified locations. DSBs are important for site-specific mutagenesis in that they stimulate the cell's natural DNA-repair processes, namely homologous recombination and non-homologous end joining. Using well-established and robust protocols, these cellular processes can be harnessed to generate precisely targeted genomic edits resulting in cell lines, including somatic cell lines, with targeted gene deletions (knockouts), integrations, or modifications. Featured lectures will provide in-depth discussion on the fundamentals of ZFNs as well as cutting edge applications for targeted gene knockout, integration and tagging. Additionally, each of the participants will conduct laboratory exercises to become proficient in various techniques for ZFN delivery, identification of mutants, and methods for quantitating ZFN efficiency. Registrants should be familiar with basic mammalian cell culture techniques, PCR and cloning.

Dates and Locations

Sigma is now able to bring the workshop to you! For more information about bringing the workshop to your institution, please contact TechnicalWorkshops@sial.com

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Lecture and Lab Topics

Targeted Genome Editing Using Zinc Finger Nucleases Lectures

• Introduction to ZFN Technology
• ZFN Design
• Genome Editing and Delivery
• Nucleofection Overview
• Double-strand Break (DSB) Repair
• Gene Knockout and Validation
• Integration Detection
• Gene Insertion and Tagging
• Donor Plasmid Design
• Dilution Cloning and Genotyping
• Quantitation of ZFN Cleavage Activity
• High-frequency ZFN-induced Gene Targeting
• Troubleshooting Exercise
• Therapeutic Application of ZFN Technology
• Creation of Transgenic Animals Using ZFN Technology

Targeted Genome Editing Using Zinc Finger Nucleases Labs

• Nucleofection
• Cel-1 Assay
• Junction PCR
• RFLP Assay

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Previous Guest Lecturers

Dana Carroll, Ph.D., Professor and Chair of Biochemistry at University of Utah
Roland Buelow, Ph.D., CEO at Open Monoclonal Technology Inc.
Morten Frödin, Ph.D., Associate Professor and Group Leader, BRIC
Bruno Reversade, Ph.D., Principal Investigator, IMB, Singapore
Detlev Arendt, Ph.D., Principal Investigator, EMBL
Dietmar Kappes, Ph.D., Director, Transgenic Mouse Facility, Fox Chase Cancer Center
David Drubin, Ph.D., Professor of Cell and Developmental Biology, University of California-Berkeley

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Feedback from Attendees

"Absolutely awesome. Everyone at Sigma is very receptive and ready to assist with all needs. I would highly recommend this workshop to anyone in the field. I am very glad that I had this opportunity." — Laia Gómez Baldó, Institute Català d'Oncologia, Spain

"An exquisitely crafted workshop with educational goals and highly recommended." — Hwei Ling Tan, National Cancer Center, Singapore

"I just wanted to thank you for the hospitality you have shown us during our stay in St. Louis. It was a very pleasant experience and it felt very good to know that there are people that not only care about the science but also about fellow human beings. The workshop was very nicely arranged, if there were any hiccups, I did not see any. The crew was also very helpful and friendly, that counts a lot for me." — Phiyani Lebea, at CSIR Biosciences, South Africa

BioU grad, Beverly Chilton talks in depth about her experiences with our workshops.

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