GENOME EDITING IN MAMMALS USING CRISPR TOOLS · 2019. 11. 21. · Ingeniería Genética y Biología...
Transcript of GENOME EDITING IN MAMMALS USING CRISPR TOOLS · 2019. 11. 21. · Ingeniería Genética y Biología...
FINAL REPORT International Workshop
GENOME EDITING IN MAMMALS USING CRISPR TOOLS
Date and venue
October 14 to 17th, 2019 Institut Pasteur de Montevideo (IPMon) Mataojo 2020, Montevideo (11400) URUGUAY
Organizers
Martina Crispo*, María Noel Meikle*, Ana Paula Mulet*, Geraldine Schlapp* Unidad de Animales Transgénicos y de Experimentación, Institut Pasteur de Montevideo, Uruguay Alejo Menchaca* Instituto de Reproducción Animal de Uruguay (IRAUy), Montevideo, Uruguay *ISTT members
Financial support
The workshop received the following grants: From IPMon: Fondo para la Convergencia Estructural del Mercosur (FOCEM) From Uruguay: Agencia Nacional de Investigación e Innovación (ANII) International: International Society for Transgenic Technologies (ISTT) and Institut Pasteur International Network (RIIP)
Invited speakers/instructors
We invited three scientific leaders in the field of CRISPR technologies, all ISTT members: Lydia Teboul – Medical Research Council ‐ Harwell Institute, United Kingdom Lluis Montoliu – Centro Nacional de Biotecnología, Spain Chris Proudfoot ‐ The Roslin Institute, University of Edinburgh, Scotland Martina Crispo and Alejo Menchaca were in charge of some lectures; Ana Paula Mulet, María Noel Meikle and Geraldine Schlapp were tutors in the informatics and practical sessions.
Workshop program and activities
This workshop offered instrumental lectures in which the uses of genome editing tools and particularly CRISPR systems were thoroughly exposed (history, principles and applications). There was a strong practical component of informatics sessions in which the design of CRISPR reagents and the use of CRISPR associated tools was explained step by step, and practical lessons in which the students had the opportunity to manipulate, microinject and electroporate mouse embryos. Also, training in diverse genotyping strategies was performed. The last day, students were asked to present their projects in order to promote discussion between students and teachers. Social events contributed to participant´s networking and exchange of experiences in a more relaxed atmosphere. The program was the following: Monday, October 14th 08:45 – 09:00 Registration 09:00 – 09:30 Teacher´s and student´s presentation 09:30 – 10:00 Unidad de Animales Transgénicos y de Experimentación – presentation of our
Unit (G. Schlapp) 10:00 ‐ 10:15 Coffee break 10:15 – 11:30 Lecture: Genome editing with programmable nucleases: general overview (L.
Montoliu) 11:30 – 12:30 Lecture: Introduction to CRISPR system (L. Teboul) 12:30 – 14:00 Lunch 14:00 – 17:00 Practical/Informatics session: step by step tutorial for the design of CRISPR
reactives (part I) (L. Teboul, C. Proudfoot, L. Montoliu, AP. Mulet) 17:00 Toast; Informal get‐together
Tuesday, October 15th 09:00 – 11:00 Practical/Informatics session: step by step tutorial for the design of CRISPR
reactives (part II) (L. Teboul, C. Proudfoot, L. Montoliu, AP. Mulet) 11:00 – 11:15 Coffee break 11:15 – 12:30 Lecture: Genome editing in embryos (L. Montoliu) 12:30 – 14:00 Lunch 14:00 – 14:30 Introduction to microinjection and electroporation of embryos (M. Crispo) 14:30 – 17:00 Practical session: Microinjection and electroporation of mouse embryos (M.
Crispo, G. Schlapp, MN. Meikle, AP. Mulet) Wednesday, October 16th 09:00 – 12:00 Theoretical‐practical session: Genotyping strategies (L. Teboul, C. Proudfoot,
L. Montoliu, AP. Mulet) 12:00 – 13:30 Lunch 13:30 – 15:30 Practical session: sequencing (L. Teboul, C. Proudfoot, L. Montoliu, AP. Mulet) 15:30 – 15:45 Coffee break 15:45 – 17:00 Round table: CRISPR troubleshooting (L. Teboul, C. Proudfoot, L. Montoliu, AP.
Mulet, M. Crispo, A. Menchaca, G. Schlapp, MN. Meikle) 18:00 ‐ 23:00 Wine cellar dinner Thursday, October 17th 09:00 – 10:00 Applications of genome editing in porcine (C. Proudfoot) 10:00 – 11:00 Applications of genome editing in sheep (A. Menchaca) 11:00 – 11:15 Coffee break 11:15 – 12:15 Alternative CRISPR applications (L. Montoliu) 12:15 ‐ 14:00 Lunch 14:00 – 17:00 Discussion of students projects & closure of the workshop
Student application and selection
Forty‐six application e‐forms were received, including students from different countries of South America but also from far countries as Turkey (ISTT member) and Egypt. Student evaluation criteria was based on: (i) clear aim of using CRISPR technologies in the applicants’ ongoing or near future projects (described in the CV, student motivation letter and supervisor’s letter); (ii) priority order was the following: PhD students; technicians; and young investigators; (iii) inclusion of candidates working in different laboratories and countries. Priority was given to people working in South America. Following these criteria, 20 students were selected, coming from Argentina (6), Brazil (3), Uruguay (5), Peru (3), Chile (1),
Paraguay (1) and Turkey (1). Only one ISTT member sent an application form (Nur Ersoy), who was selected to participate as committed with the ISTT funding grant. Selected students were the following:
Surname and name Institution City and country
Alberio Virgilia Universidad de Buenos Aires Buenos Aires, Argentina
Basika Tatiana Institut Pasteur de Montevideo Montevideo, Uruguay
Benitez Riquelme Diego University de Concepcion Concepcion, Chile
Caporale Alfredo Instituto de Investigaciones en Ingeniería Genética y Biología
Molecular Buenos Aires, Argentina
De Souza Marcela Institut Pasteur de Montevideo Montevideo, Uruguay
Ersoy Nur Istanbul University‐Cerrahpasa,
Veterinary Faculty Istanbul, Turkey
Ferreira Jessica Universidad Federal Rural de Rio
de Janeiro Rio de Janeiro, Brazil
Giletti Andrea Facultad de Química, Universidad
de la República Montevideo, Uruguay
Isaja Luciana Instituto Fleni Buenos Aires, Argentina
La Motta Gastón Facultad de Agronomía,
Universidad de Buenos Aires Buenos Aires, Argentina
Lasalle André Facultad de Ciencias, Universidad
de la República Montevideo, Uruguay
Laymito Lina Rocio Universidad de San Martin de
Porres Lima, Peru
Li Calzi Marco Institut Pasteur de Montevideo Montevideo, Uruguay
Luque Guillermina Instituto de Biología y Medicina
Experimental Buenos Aires, Argentina
Marks Maria Paula Instituto de Biología y Medicina
Experimental Buenos Aires, Argentina
Melo Calero Omar Universidad Peruana Cayetano
Heredia Lima, Peru
Méndez Alejandro Universidad Nacional de Asunción, Facultad de Ciencias Exactas y
Naturales Asunción, Paraguay
Pereira Talita Pontificia Universidade Catolica do
Rio Grande do Sul Porto Alegre, Brazil
Ponce‐Salazar Miguel Vivanco International Lima, Peru
Silva Alexandre Instituto Oswaldo Cruz Rio de Janeiro, Brazil
Organization
The Institut Pasteur de Montevideo is located at short distance from hotel infrastructure in Montevideo. Teachers and students were accommodated in the same hotel, which was important to foster interaction. A transfer bus ensured daily hotel/institute/hotel transport. The institute has designated laboratories for teaching purposes, for both practical and informatics sessions. Secretarial and technical assistance is available, both for organizational logistics, as well as for informatics system management. Furthermore, the institute and our
unit count with good previous experience in organizing theoretical and practical courses, which resulted in a very smooth flow, with no major problems. The Secretary of the IP Montevideo took care of every non‐academic detail, from scheduling flight arrivals and departures to dealing with food, accommodation and travel providers.
Global appreciation of the training
The main objective of this workshop was to update scientist and technicians, working in the generation of GM animal models, with the latest available technology regarding CRISPR/Cas. In addition, the generation of a regional network in this field was especially encouraged. Although there was a good balance between theoretical and practical lessons, hands‐on training was a key component. Since the success of a new GM animal model depends mostly on the designing of the CRISPR tools, special emphasis was given to informatics sessions. Moreover, during practical lessons (and coffee breaks), students discussed their own projects with the teachers, which was a very valuable aspect of this event. After the workshop, most of the students were capable of: ‐ Choosing a suitable scientific approach ‐ Designing the appropriate CRISPR tools for their projects ‐ Delivering the CRISPR system into mammalian zygotes by microinjection or electroporation ‐ Analyzing mutations in vitro and in silico ‐ Discussing with the teachers about their own projects The Workshop was entirely dictated in English. A hands‐on manual was prepared by the organizers. The participants were mostly advanced Master or PhD students, post‐doctoral fellows, technicians and young research scientists. Students had the opportunity to present their own research work, in a 10 minute powerpoint presentation. Diverse on‐going and future projects were presented and discussion was encouraged. At the end of the workshop, students were asked to fill out an anonymous survey form of the workshop, to evaluate different aspects of this event, which resulted very valuable for the organizing team. Survey results are attached to this report. Finally, an exam was available for those students who needed the credits to be presented in their postgraduate programs.
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