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Genome Editing Research Group (GERG)

Mission

The mission of the Genome Editing Research Group is to collect and disseminate the fast-evolving knowledge in designer nuclease technologies, to undertake pilot research projects and to provide/publish the streamlined processes that could be used as guidelines by the interested Core Facilities to initiate Genome Editing technologies as services in their labs.

Current Membership

Elizabeth Sergison (Chair) - Dartmouth College

Maureen Regan (Co-Chair)- University of Illinois at Chicago 

Kym Delventhal - Stowers Institute 

Channabasavaiah Gurumurthy  - University of Nebraska Medical Center

Eric Kmiec - Gene Editing Institute

Shondra M. Pruett-Miller - St. Jude Children's Research Hospital

Timothy J. Dahlem - Recursion Pharmaceuticals

Gerald MarsischkyIndependent Consultant

Kevin Knudtson (EB Liaison) - University of Iowa

 

Studies

GERG 2019 Study

 

Recent advances in genome engineering are allowing scientists to better understand biology by precisely deleting, editing, or tagging genomic DNA. The clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated (Cas) system was first used to edit mammalian cells in 2013 and has grown in popularity ever since. Multiple guideRNA and Cas9 reagent formats can be used for editing cells. In this study, we compared three popular methods: 1. a plasmid expressing both the guideRNA and Cas9, 2. Cas9 protein combined with a synthetic single guideRNA, and 3. Cas9 combined with a 2-part guideRNA. In addition, the CRISPR/Cas system can be delivered to cells via lipofection or nucleofection transfection methods. This study aims to compare the efficiency of gene editing outcomes at 3 different genomic targets, 3 unique guideRNA reagent formats, and 2 delivery method across multiple labs. For the 2018 GERG study, the group performed a pilot study and found that the results varied considerably across the 4 sites. Three possible sources of the variation are: 1. researchers had different levels of experience with the different methods 2. the provided protocols (from the companies) were challenging to understand, and 3. each researcher only performed one replicate. In 2019, we wrote a standard protocol and repeated the experiments multiple times to more accurately evaluate the reproducibility of these methods. Determining which CRISPR reagent format is the most reproducible and has the highest gene editing outcomes will be beneficial for core facilities or research labs getting started with genome editing.

Results were presented in a poster at ABRF 2020:  GERG Study 2019-2020: Reproducibility of indel formation rates by comparing guideRNA format and delivery method

 

 

GERG 2018 Reproducibility Study

In 2018, GERG initiated a study to evaluate the reproducibility of indel formation rates by comparing guideRNA format and cell delivery methods across multiple labs.  Various configurations of guideRNA and Cas9 components can be used for editing cells.  A few options include: a plasmid expressing both the guideRNA and Cas9, Cas9 protein combined with a synthetic single guideRNA, and Cas9 combined with a synthetic 2-part guideRNA.  In addition, delivering these components to cells can be done using lipofection or nucleofection transfection methods.  In the GERG 2017 survey (CRISPR/Cas9 Methods:  Preferences from the Field), plasmid format and lipofection delivery were favored among cell culture users.  Meanwhile, RNP format for the guideRNA and Cas9 is gaining in popularity in combination with nucleofection delivery.  This study aims to evaluate cutting efficiency at 3 different guideRNA targets based on the guideRNA format and delivery method across multiple labs.  Determining which method or format is the most reproducible will be beneficial.  Core facilities or research labs getting started with genome editing could use these results as a benchmark for optimizing their own protocols.

Results were presented in a presentation at ABRF 2019:  Reproducibility of indel formation rates by comparing guideRNA format and delivery method

Results were also presented in a poster at ABRF 2019:  Reproducibility of indel formation rates by comparing guideRNA format and delivery method

GERG Survey 2017

The Genome Editing Research Group surveyed users of the CRISPR/Cas9 technology to help establish an understanding of preferred methods being used.  As new core facilities are being formed to support the CRISPR/Cas9 technology, or existing cores have adapted their services to fit the technology into their workflows, considering what other cores are using is important.   Questions regarding preferred guideRNA design tools, format of reagents, mutation analysis methods, and other relevant topics were included. 

The collected data can be viewed here:  https://www.surveymonkey.com/results/SM-ZBM5L6768/

Results were summarized in a poster and presented at ABRF 2018:   CRISPR/Cas9 Methods:  Preferences from the Field 

 

Membership History

 

    

 

Name

Organization (during membership)

Member

Chair

Co-Chair

Channabasavaiah Gurumurthy

University of Nebraska Medical Center

03-15

03-15 to 04-17

 

Kym Delventhal

Stowers Institute for Medical Research

03-15

04-17 to 05-19

 

TJ Craddick

Georgia Institute of Technology, Emory University

03-15 to 12-16

 

 

Vittorio Sebastiano

Standford University

05-15 to 12-16

 

 

Eric Kmiec

Gene Editing Institute

09-15

 

 

Shondra Pruett-Miller

Washington University, St. Jude Children's Research Hospital

12-15

 

 

Timothy Dahlem

University of Utah

07-16

 

 

Elizabeth Sergison

Dartmouth College

05-17

05-19

04-18 to 05-19

Gerald Marsischky

Independent Consultant

02-17

 

 

Maureen Regan

University of Illinois at Chicago

09-18

 

05-19

 

 

 

 

 

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