Customer Education

From Genotype to Phenotype: Emerging Technologies for CRISPR Genome Editing and Molecular and Cellular Assays

Seminar Title: From Genotype to Phenotype: Emerging Technologies for CRISPR Genome Editing and Molecular and Cellular Assays

Date: Wednesday, October 25, 2017

Time: 2:00 PM – 4:00 PM (short break between talks)
Registration and Check-in: 1:45 PM

Location: Johns Hopkins University
Miller Research Building (MRB) G01



 Lecture 1: Recent Advances in CRISPR/Cas9 Genome Editing Technology: From eSpCas9
 to Whole-Genome Screening

Time: 2:00 PM – 2:50 PM
 

 Abstract

Genome editing has enabled the investigation of gene function and target identification, and is helping researchers develop potential therapies against a wide range of diseases. MilliporeSigma has been an industry leader in genome editing technology since 2002 when it introduced the TargeTron system for gene editing in prokaryotes. Five years later, it became the first company to commercialize eukaryotic genome editing technology in the form of Zinc Finger Nucleases (ZFNs). The company was at the forefront of the CRISPR revolution in 2012, and continues to develop cutting-edge genome and epigenome editing tools with ever greater target specificity and robust cutting activity.

After an introduction to CRISPR/Cas9 technology, this presentation will highlight the latest developments in the field of genome editing. We will discuss strategies to enhance homology-directed repair (HDR) to facilitate gene insertion or swap, including Cas9-Geminin. We will describe enhanced-specificity Cas9 (eSpCas9), which is a rationally engineered version of wild-type SpCas9 with significantly fewer off-target effects. We also will illustrate the proxy CRISPR targeting strategy, which makes CRISPR gene editing more efficient, flexible, and specific. We will explain epigenetic applications of CRISPR technology, including dCas9-p300 and CRISPR-SAM activators. We also will highlight pooled and arrayed CRISPR libraries, such as the widely-acclaimed Sanger LentiCRISPR array, which allows whole-genome or gene family screening. The presentation will end with a brief overview of other emerging technologies available from MilliporeSigma.

Topics that will be discussed include:

  1. CRISPR Background & Mechanism
  2. Recent CRISPR Innovations
       a. HDR enhancement
       b. SygRNA & eSpCas9
       c. Proxy CRISPR
  3. Exciting CRISPR Applications
       a. Epigenetic activators
       b. Whole genome lentiviral arrays & pools
  4. Emerging Technologies from MilliporeSigma

 

Speaker Bio
M. Zulfiquer Hossain, PhD
Research Technology Specialist
MilliporeSigma
Dr. Hossain received his BS in Biology and Biochemistry from Brandeis University and his PhD in Molecular Medicine from the University of Maryland under the Graduate Partnerships Program (GPP) with the National Institutes of Health (NIH). He has over 10 years of biomedical research experience, both as a pre-doctoral fellow at the NIH and as a post-doctoral fellow at the Johns Hopkins University, where he identified novel DNA-damaging polyphenols in foods and flavorings. He brings extensive knowledge of molecular, cancer and stem cell biology, epigenetics and immunology, as well as teaching experience as an Associate Professor at BRAC University. As a Research Technology Specialist with MilliporeSigma, Dr. Hossain supports research projects that use cutting-edge functional genomics technologies.

 Lecture 2: What Have I Done?!: How to Determine the Effects of Gene Editing on
 Cellular Phenotype

Time: 3:00 PM – 3:50 PM
 

 Abstract

The use of innovative gene editing technologies, such as CRISPR/Cas9, continues to become more common in a variety of research areas, including oncology and infectious diseases. Manipulating characteristics of genomic DNA or RNA expression levels can have profound effects on cellular protein levels, and, thus, viability and overall phenotype of the targeted cells. Therefore, the downstream effects of any genetic manipulation must be thoroughly assessed and understood in order for researchers to accurately interpret resultant experimental data.  In this presentation, we will discuss how protein-based analysis using flow cytometry and quantitative, multiplexing protein concentration measurements can be easily integrated into any study to achieve phenotyping goals. We will highlight the combined use of CRISPR/Cas9, Guava flow cytometers, and Milliplex protein multiplexing assays from recently published literature and will be available to discuss specific study-design questions.

Topics that will be discussed include:

  1. Overview of Guava flow cytometry technology and uses for analyzing cell health
  2. Overview of Luminex xMAP® technology and Milliplex®MAP multiplexing assay panels

 

Speaker Bio
Karen R. Tamul, MS, MT(ASCP)SI
Field Application Scientist
MilliporeSigma
Ms. Tamul has extensive experience in Immunology with a focus on flow cytometry, gained through a varied career that includes clinical hospital and reference laboratories, proficiency testing services, and sales and applications positions with a number of leading manufacturers in the field for both instruments and reagents. She has served with MilliporeSigma as a Field Application Scientist for seven years.

 

Speaker Bio
Jason M. Rosenzweig, PhD
Biomarker and Cytometry Specialist
MilliporeSigma
Dr. Rosenzweig received a PhD in Cellular and Molecular Medicine from Johns Hopkins University. He has extensive experience in immunology, biomarker discovery, flow cytometry, and proteomics and provides solutions to enable and enhance life science research.



For more information, please contact technicalworkshops@sial.com or:

Michelle Laird
Phone: 301-395-6719
Email: michelle.laird@sial.com