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Advanced Gene Editing with CRISPR — 3 Day Training Workshop

December 7, 2022 @ 9:00 AM December 9, 2022 @ 5:00 PM


Advanced Gene Editing with CRISPR is a 3-day hands-on workshop designed for researchers with entry level experience in CRISPR/Cas9 technology seeking a more in-depth immersion in state-of-art CRISPR applications such as working with sgRNA functional libraries, disease modeling using CRISPR, successful employing of CRISPR technology in complex experimental systems.

Note: This workshop is ideal for those who have already participated in the Bio-Trac “Gene Editing with CRISPR” workshop.  Attendees should be experienced in common technologies of manipulating cell cultures and isolation / characterization of macromolecules, such as cellular proteins and genomic DNA. Prior experience in cell line transfections, polymerase chain reaction and protein gel-electrophoresis/Western blotting technologies will be beneficial.

Lectures Topics

  • Advanced Applications of CRISPR Technology I: Precllinical Modeling of Diseases
  • Studying Vertebrate Development and Cell Signalling Using CRISPR Genome Editing in Non-Murine Models
  • Advanced Applications of CRISPR Technologies II:  Therapeutic and Diagnostic Aspects
  • Advanced Applications of CRISPR Technologies III: Construction and Application of CRISPR Libraries for Functional Screening and Gene Identification Purposes
  • CRISPR Technology – Synthego
  • Drug Target Identification and Drug Screening In vitro Using CRISPR Methodology
  • Strategies for Delivering CRISPR Reagents In Vitro and In Vivo: Efficiency vs. Immunogeicity vs. Toxicity vs. Biosafety Considerations
  • CRISPR Technology: Invitrogen Attune NxT Flow Cytometry – Off Target Detection
  • Evaluation of Endotoxin and Other Bacterial Contaminants in Cas9 Preparations
  • Discussion: Experimental Design for Successful CRISPR/Cas9 Project Aimed at Precise Genome Editing Pursuing HDR Outcomes
  • Discussion: Next Steps and Further CRISPR Applications

Laboratory 1:

  • Assembly of CRISPR/sgRNA RNP complexes for cell transfections
  • Preparation of Cells for In Vitro Transfection of RNP Complexes
  • Transfection of Cells with RNP Complexes
  • Collection of Set I (Day I) of Transfected Cells and Genomic DNA Isolation
  • Collection of Set II (Day 2) of Transfected Cells and Genomic DNA Isolation
  • PCR Amplification of Targeted Region and RLFP Confirmation of HDR events
  • Fluorescent Sorting Analyses of HDR Efficiency

Laboratory II

  • Preparation of Bacterial Cultures for Prokaryotic Expression of Cas9 Enzymes
  • Induction of Bacterial Cultures for Recombinant Cas9 Proteins Expression
  • Preparation of Protein Lyzates from Cas9 Expressing Bacterial Cultures
  • Isolation of 6xHis-Cas9 Proteins Using Ni-NTA Affinity Columns
  • PAGE Gel Electrophoresis of Bacterial Lyzates and Affinity Purified Cas9 Proteins
  • Setting Western Blot Transfer of PAGE Separated Proteins
  • Visualizing Recombinant Cas9 Proteins on Westent Blots by anti-6xHis Antibodies
  • Estimating the Yield of Recombinant Cas9 Isoforms 


December 7, 2022 @ 9:00 AM
December 9, 2022 @ 5:00 PM
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“As always, Bio-Trac not only met my expectations, but exceeded them! I was immensely impressed with the class – both the lab work and lectures! I will absolutely be taking more classes with BioTrac – their classes should be required for all graduates entering the field of science. Easy to understand & follow, yet the classes get to the core of the discussed principle. I loved learning about CRISPR and would highly recommend this class to anyone in the field!”

Jennie Rowell PhD, RN
Assistant Professor,
The Ohio State University

“This workshop provided and excellent opportunity to further my understanding of the CRISPR technology. Dr. Kozlov presented clear and engaging lectures and hand-on lab time that gave me better understanding of the evolving CRISPR field and an appreciation for the lab techniques involved. The best part of the workshop were the ample question and answer periods that allowed me to explore my specific research concerns about CRISPR with experts in the field.”

Robert Yasuda, PhD
Assistant Professor of Pharmacology, Georgetown University

“This course is awesome! I not only learned a lot about very important up-to-date CRISPR/CAS9 technology, I also became very excited to bring this information back to Xavier to share it with faculty. Hopefully, I can integrate CRISPR/CAS9 as a CORE offering.”

Melissa Bratton
Cellular and Molecular Research Scientist
Xavier University of Louisiana


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20200 Observation Drive, Room 151/152
Germantown, MD 20876 United States
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