Dec 7-9, 2022


Germantown, MD

The Bioscience Education Center


1 Hour Lunch Break

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.

Lecture and Hands-on Interactive Training
Team taught by active researchers
thumbnail drive
Thumbnail drive with Lectures and Workshop material
Space limited to 24 participants
Registration Fee: $895

Course Director

Course Director

Serguei Kozlov, PhD, MBA, PM
Team Leader, Principal Scientist and Program Manager, Center for Advanced Preclinical Research, Leidos Biomedical Research, Inc.

  • 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 

Serguei Kozlov, PhD, MBA, PM

Dr. Kozlov received his Master of Science diploma in applied mathematics and theoretical physics from Moscow Institute of Physics and Technology in Russia, subsequently earning a PhD degree in molecular genetics and neurobiology from the Institute of Biochemistry, University of Zurich, Switzerland. He subsequently received a Master of Business Administration degree from Robert Smith School of Business, University of Maryland and also holds several other professional certifications and credentials.

Dr. Kozlov joined the National Cancer Institute in 1999 as a postdoctoral fellow in Cancer and Developmental Biology lab where he was promoted in 2003 to senior research fellow and subsequently in 2007 to the staff scientist appointment. In 2008 he joined SAIC Inc. (later renamed to Leidos BMR, Inc.) where he established Center for Advanced Preclinical Research – a translational resource providing interdisciplinary support and expertise in establishing next-generation animal cancer models, developing technologies for advanced manipulation of mammalian genomes for the purpose of preclinical modeling, and directing early stage drug development projects to accelerate clinical introduction of promising efficacious cancer therapeutics.

The Bioscience Education Center

Montgomery College
20200 Observation Drive
Germantown, MD 20876