14th Transgenic Technology Meeting

October 1-4, 2017 Salt Lake City,
Utah, USA Snowbird Resort

Workshops

Workshop registration is now closed.  This is a hands-on workshop that will have some overlap with the mouse workshop running simultaneously regarding CRISPR design and synthesis of reagents.

Participation is subject to a selection process. To be eligible, you must upload a short CV, letter of motivation (max 200 words), and a short letter of support during conference registration.   Selected participants will be notified by email and charged the additional registration fee.

Note that Workshop 1 and Workshop 2 will have some combined lectures and sessions in addition to breakout hands-on sessions.

Suggested housing for the workshop is The University Guest House, the on-campus hotel at the University of Utah. A standard room will be $112.00/night plus tax. The workshops will be held in a nearby campus building, a short 10 minute walk from the hotel.

Workshop 1: Basics and latest trends in CRISPR mouse genome editing technologies                                

Sponsored by:charles_river_logo

The workshop will include lectures and demonstrations of CRISPR mouse genome engineering experiments using the advanced techniques such as Easi-CRISPR (Efficient additions with ssDNA inserts CRISPR) and GONAD (Genome editing via Oviductal Nucleic Acids Delivery). The participants will have opportunity to closely interact with the instructors and perform hands-on experiments such as designing CRISPR genome engineering projects, synthesis of CRISPR and donors reagents for microinjection and electroporation (GONAD) experiments, and either watch or perform microinjection and/or GONAD procedures.

  • Organized by Channabasavaiah Gurumurthy (Guru)
    • Assisted by Rolen Quadros, Donald Harms, Masato Ohtsuka, Masahiro Sato, Hiromi Miura, Paul Bonthius, Rick Focht, Tamowski lab, Mansour lab

Date: September 28, 1pm – October 1, 11am

Location: University of Utah

Participation Fee: $350

Thursday, September 28, 2017

1:00 pmIntroduction, orientation (Susan Tamowski)
1:15 pmDavid Grunwald lecture, “An introduction to engineered nucleases and their applications for: modifying gene expression, targeted mutagenesis, homology-directed repair, and homologous recombination”; ”Strategies for genome modification in the zebrafish”
2:00 pmGuru lecture, “Overview of mouse genome engineering using CRISPR/Cas9 system”
2:45 pmStudent short introductions/research focus/experience; preferably present 2-3 slides each, or self-introduction (without slides).
3:45 pmTour of workshop rooms and facilities
4:00 pmLab session: set up PCR reaction

Friday, September 29, 2017

8:00 - 9:30 amLab session (purification of PCR products and setting up RNA synthesis)
9:30 - 10:15 amBasics and advances in CRISPR mouse genome engineering methods (Guru)
10:15 - 11:00 amGONAD; a novel genome engineering method (Masato)
11:00 - 12:00 pmExamples of designing conditional and knock-in alleles (Guru)
12:00pmLUNCH
1:00pmTimothy Dahlem lecture, “Design and construction of gRNA’s”
2:00pmComputer lab session: Hands-on instruction aimed at designing gRNAs to a gene(s) of your interest
3:00 - 4:00 pmLab session (purification of RNA and setting up RT reaction)
4:00 - 5:00 pmGONAD demonstration (Masato, Masahiro, Hiromi)
5:00 - 6:00 pmGONAD practice (Group A students)

Saturday, September 30, 2017

8:30 amLab session (purification of ssDNA and gel analysis)
9:30 amPreparing Easi-CRISPR microinjection mix (presentation by Rolen)
10:00 amLab session (preparation of Easi-CRISPR microinjection mix)
10:30 - 12:00 pmLab session (demonstration of thawing zygotes and microinjection), Don, Rick & Tamowski lab personnel
12:00pmLUNCH
1:00pmLab session (harvesting of zygotes from the GONAD treated females and observation of fluorescence)
2:00 - 3:45 pmMicroinjection practice (students of all four groups)
3:45 - 6:00 pmGONAD practice (Groups B, C and D students)

Sunday, October 1, 2017

9:00 amLook at injected embryos, harvesting of zygotes from the GONAD treated females and observation of fluorescence
10:00 amGroup discussion on molecular biology and GONAD results
10:30 amDesigning student’s Easi-CRISPR projects
Closing comments

Partial Workshop 1 Sponsor: BEXLogo300ppi_95799

Some molecular biology reagents were generously donated by:

neb_logo_cmykIDT_Logo16_RGB

Workshop 2: An Introduction to Genome Modification Methods in Zebrafish

This workshop is intended to be a conceptual and practical introduction to current methods used for editing the zebrafish genome. We will discuss principles of using engineered nucleases to induce targeted mutations in zebrafish and to generate precisely edited alleles by homologous recombination. Students will get hands-on instruction to design Crispr/Cas9 targeting tools, to inject fertilized eggs with Crispr/Cas9 reagents, and to detect induced mutations by phenotypic screening and by analysis of genomic DNA.  Principles of phenotypic analysis and the use of transgene reporters will also be discussed.

  • Organized by Timothy Dahlem, Richard Dorsky, David Jonah Grunwald and Kristen Kwan

    • Additional Course Instructors: Susan Tamowski and Channabasavaiah Gurumurthy
    • Course Teaching Assistants: Maurine Hobbs, Chase Bryan, Hannah Gordon, Sarah Lusk, Dana Shaw, Deeptha Vasudevan, Alex Chagovetz

Date: September 28, 1pm – October 1, 11am

Location: University of Utah

Participation Fee: $350

Thursday, September 28, 2017

1:00 pmSusan Tamowski: Introduction to workshops
1:15 pmDavid Grunwald lecture: An introduction to engineered nucleases and their applications for: modifying gene expression, targeted mutagenesis, Homology- Directed Repair, and Homologous Recombination"; “Strategies for genome modification in the zebrafish”
2:00 pmChannabasavaiah Gurumurthy lecture: “Overview of mouse genome engineering using CRISPR/Cas9 system”
2:45 pmStudent short introductions/research focus/experience; preferably present 2-3 slides each, or self-introduction (without slides).
3:45 pmTour of workshop rooms and facilities
4:00 pmCourse instructors: “An overview of zebrafish development and the experiments of the workshop”
Friday, September 29, 2017
8:30amKristen Kwan lecture, “Overview of spawning, egg collection, and microinjection”
9:00amLab session: Egg collection and microinjection of zebrafish embryos
Experiments: injections of GFP mRNA albino and rx3 CRISPR/Cas9 complexes
12:00pmLUNCH
1:00pmTimothy Dahlem lecture, “Design and construction of gRNA’s”
2:00pmComputer lab session: Hands-on instruction aimed at designing gRNAs to a gene(s) of your interest
3:00pmObserve embryo development; care of embryos; look for GFP expression; set up matings
4:30pmKristen Kwan lecture: “Generation and use of tissue-specific reporters”
Richard Dorsky lecture: “Approaches to phenotypic analyses
Saturday, September 30, 2017
9:00amLab session: Injection of embryos
Experiments: injections of GFP mRNA albino and rx3 CRISPR/Cas9 complexes
12:00pmLUNCH
1:00pmTimothy Dahlem lecture: “Detection of newly induced mutations”
1:30pmTimothy Dahlem hands-on session using HRMA to detect induced mutations
4:00pmObserve embryos development; previous day’s injections; look for GRP expression; look for induced mutant phenotypes
Sunday, October 1, 2017
9:00amObserve embryo development; look for GRP expression; look for induced mutant phenotypes Closing comments