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
2:30 - 3:45 pmLab session (purification of RNA and setting up RT reaction)
3:45 - 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.

Workshop 2: An Introduction to Genome Modification Methods in Zebrafish

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

  • Additional Course Instructors: Susan Tamowski and Channabasavaiah Gurumurthy
  • Course Teaching Assistants: Chase Bryan, Dr. 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