ANAT2341 Lab 4: Difference between revisions

Latest revision as of 10:52, 24 June 2019

1. QUIZ

2. Guest Lecturer - Dr Fabien Delerue

Dr Fabien Delerue

Manipulating the mouse embryo: from ES Cells to genome editing

Doctor Fabien Delerue is the Head of the Genome Editing at Macquarie (GEM), a transgenic core developed in joint venture by the Dementia Research Centre (DRC) and the Macquarie Animal Research Services (MARS). Fabien graduated from the University of Bordeaux in France and received his PhD from the University of Sydney in 2013. Fabien gained his first research experience in the preclinical CNS Centre of Hoffmann-La Roche Ltd (Basel, Switzerland) where he became an expert in in vivo activities (i.e. animal models of diseases). In 2013, Fabien moved to the University of New South Wales, where he became Lecturer and managed the Transgenic Animal Unit. His research focused on developing new gene transfer techniques particularly applicable to dementia and rare genetic diseases. In 2018 he became Senior Lecturer and joined Macquarie University to head the GEM transgenic facility and develop in parallel a research program on rare genetic disorders. Research interests

Fabien’s contribution to the field of transgenesis and genome editing is internationally recognized. Fabien’s career spans over 20 years practice in assisted reproduction and generation of animal models of diseases. Current projects include:

- Optimization of gene transfer methods for the generation of transgenic animal models

- Development of new assisted reproductive techniques

- Discovery and engineering of new endonucleases

- Investigating the pathomechanisms of FoxG1 syndrome

Lecture slides: File:Delerue.pdf

References

Delerue F & Ittner LM. Genome Editing in Mice Using CRISPR/Cas9: Achievements and Prospects Cloning and Transgenesis 2015, 4:135 doi:10.4172/2168-9849.1000135 [1]

"Animal models are a powerful tool to understand the mechanisms underlying physiological and pathological processes in vivo. To date, mice remain the species most commonly used for genetic manipulation. The recent development of engineered endonucleases such as Zinc Finger Nucleases (ZFN), Transcription activator-like effector nucleases (TALEN), and the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9) offered easy, flexible, and fast alternative to ES-Cell based gene targeting. Thanks to multiple advantages, the CRISPR system superseded its predecessors and became a popular method for genome editing. Here, we review the latest techniques to apply CRISPR editing to the mouse genome, and emphasize on the current methods used in transgenic laboratories and subsequent achievements in mice."

Ke YD, van Hummel A, Stevens CH, Gladbach A, Ippati S, Bi M, Lee WS, Krüger S, van der Hoven J, Volkerling A, Bongers A, Halliday G, Haass NK, Kiernan M, Delerue F, Ittner LM. Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS. Acta Neuropathologica 2015 Nov;130(5):661-78 PubMed 26437864 [2]

Ittner A, Chua SW, Bertz J, Volkerling A, van der Hoven J, Gladbach A, Przybyla M, Bi M, van Hummel A, Stevens CH, Ippati S, Suh LS, Macmillan A, Sutherland G, Kril JJ, Silva APG, Mackay J, Poljak A, Delerue F, Ke YD, Ittner LM. Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer’s mice Science 2016 Nov;354 (6314):904-908 doi: 10.1126/science.aah6205 [3]

Recommended Literature


Manipulating the Mouse Embryo: A Laboratory Manual (Fourth Edition) By Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy, & Andras Nagy.	Reproductive Engineering techniques in mice: Technical manual Prof Naomi Nakagata, Center for Animal Resources and Development, Kumamoto University, Japan

External Links

External Links Notice - The dynamic nature of the internet may mean that some of these listed links may no longer function. If the link no longer works search the web with the link text or name. Links to any external commercial sites are provided for information purposes only and should never be considered an endorsement. UNSW Embryology is provided as an educational resource with no clinical information or commercial affiliation.

NCBI Search - NCBI databases - CRISPR | PubMed CRISPR | PubMed Centrap CRISPR
JoVE - [4]

<html5media height="460" width="640">https://www.youtube.com/watch?v=16g6fEup2pQ</html5media>

@@ Line 1: / Line 1: @@
-{{ANAT2341Lab4}}
+== 1. QUIZ ==
-=Endoderm Development=
+== 2. Guest Lecturer - Dr Fabien Delerue ==
+{|
+| width=185px|[[File:Fabien deleRue profile photo.jpg|180px]]
+Dr Fabien Delerue
+| '''Manipulating the mouse embryo: from ES Cells to genome editing'''
+<br>
+Doctor Fabien Delerue is the Head of the Genome Editing at Macquarie (GEM), a transgenic core developed in joint venture by the Dementia Research Centre (DRC) and the Macquarie Animal Research Services (MARS). Fabien graduated from the University of Bordeaux in France and received his PhD from the University of Sydney in 2013. Fabien gained his first research experience in the preclinical CNS Centre of Hoffmann-La Roche Ltd (Basel, Switzerland) where he became an expert in in vivo activities (i.e. animal models of diseases). In 2013, Fabien moved to the University of New South Wales, where he became Lecturer and managed the Transgenic Animal Unit. His research focused on developing new gene transfer techniques particularly applicable to dementia and rare genetic diseases. In 2018 he became Senior Lecturer and joined Macquarie University to head the GEM transgenic facility and develop in parallel a research program on rare genetic disorders.
+Research interests
-== 1. QUIZ ==
+Fabien’s contribution to the field of transgenesis and genome editing is internationally recognized. Fabien’s career spans over 20 years practice in assisted reproduction and generation of animal models of diseases. Current projects include:
+-      Optimization of gene transfer methods for the generation of transgenic animal models
+-      Development of new assisted reproductive techniques
+-      Discovery and engineering of new endonucleases
+-      Investigating the pathomechanisms of FoxG1 syndrome
+|}
+'''Lecture slides:''' [[File:Delerue.pdf]]
+===References===
+Delerue F & Ittner LM. Genome Editing in Mice Using CRISPR/Cas9: Achievements and Prospects
+Cloning and Transgenesis 2015, 4:135 doi:10.4172/2168-9849.1000135 [http://www.omicsgroup.org/journals/genome-editing-in-mice-using-crisprcas9-achievements-and-prospects-2168-9849-1000135.php?aid=50992]
-==2. Guest Lecturer Dr. Nicolas Fossat (CMRI) - Endoderm, RNA biology and gut organoids ==
+:"Animal models are a powerful tool to understand the mechanisms underlying physiological and pathological processes in vivo. To date, mice remain the species most commonly used for genetic manipulation. The recent development of engineered endonucleases such as Zinc Finger Nucleases (ZFN), Transcription activator-like effector nucleases  (TALEN), and the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9) offered easy, flexible, and fast alternative to ES-Cell based gene targeting. Thanks to multiple advantages, the CRISPR system superseded its predecessors and became a popular method for genome editing. Here, we review the latest techniques to apply CRISPR editing to the mouse genome, and emphasize on the current methods used in transgenic laboratories and subsequent achievements in mice."
-[[File:Nicolas_Fossat_profile_photo.jpg]]
+Ke YD, van Hummel A, Stevens CH, Gladbach A, Ippati S, Bi M, Lee WS, Krüger S, van der Hoven J, Volkerling A, Bongers A, Halliday G, Haass NK, Kiernan M, Delerue F, Ittner LM. Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS. Acta Neuropathologica 2015 Nov;130(5):661-78 PubMed 26437864 [https://dx.doi.org/10.1007/s00401-015-1486-0]
-Doctor Nicolas Fossat is a Research Fellow in Professor Patrick Tam’s laboratory, in the Embryology Unit, Children’s Medical Research Institute.
+Ittner A, Chua SW, Bertz J, Volkerling A, van der Hoven J, Gladbach A, Przybyla M, Bi M, van Hummel A, Stevens CH, Ippati S, Suh LS, Macmillan A, Sutherland G, Kril JJ, Silva APG, Mackay J, Poljak A, Delerue F,  Ke YD, Ittner LM. Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer’s mice
+Science 2016 Nov;354 (6314):904-908 doi: 10.1126/science.aah6205 [http://science.sciencemag.org/content/354/6314/904.full]
-Dr Fossat completed his PhD in 2005 at the Ecole Normale Superieure of Lyon in France. During this period, he studied the homeodomain transcription factor Otx2 which is coded by one of the few master genes essential for the formation of the head. Notably, using the latest tools of molecular biology, he developed a novel genetic approach called “conditional self-knockout” allowing the inactivation of the Otx2 gene at anytime and anywhere in the mouse. His background is a strong understanding of and an easy ability to manipulate the mouse embryo genome. In 2007, he joined Professor Patrick Tam’s team. Professor Tam is a pioneer in mouse embryo manipulation. In his lab, he pursues his understanding of the formation of the head. He focuses on the intersection of transcriptional and signalling activities by studying the interaction of another important actor of the formation of the head, the master gene Lhx1, and a fundamental pathway, the WNT signalling pathway which is crucial in every part of the body for the control of cell proliferation and cell differentiation. In addition, he works on the foregut endoderm which is also important for the formation of the head as well as for the establishment of organs involved in secretory or absorptive functions, such as the thyroid, pancreas and the liver. With the ultimate goal to cure diseases affecting these organs such as hypo/hyper-thyroidism or diabetes, he has started to study the function of an unknown gene that Patrick Tam’s lab has identified as specifically active in the foregut endoderm. This new gene codes a RNA-binding protein which has led me to also take a great interest in RNA and miRNA functions.
+===Recommended Literature===
+{|
+| [[File:Manipulating_the_Mouse_Embryo.jpg|200px]]
+| | [[File:Reproductive Engineering Techniques.jpg|200px]]
+|-
+| [http://cshlpress.com/default.tpl?cart=147944968860031853&action=full&--eqskudatarq=982 Manipulating the Mouse Embryo: A Laboratory Manual (Fourth Edition)] By Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy, & Andras Nagy.
+| [http://card.medic.kumamoto-u.ac.jp/card/english/sigen/manual/onlinemanual.html Reproductive Engineering techniques in mice: Technical manual] Prof Naomi Nakagata, Center for Animal Resources and Development, Kumamoto University, Japan
+|}
-==References==
+===External Links===
+{{External Links}}
+* NCBI Search - [http://www.ncbi.nlm.nih.gov/gquery/?term=CrispR NCBI databases - CRISPR] |  [http://www.ncbi.nlm.nih.gov/pubmed/?term=CrispR PubMed CRISPR] | [http://www.ncbi.nlm.nih.gov/pmc/?term=CrispR PubMed Centrap CRISPR]
+* JoVE - [https://www.jove.com/video/55765/generation-genetically-modified-mice-through-microinjection]
-{{2016ANAT2341 footer}}
+<html5media height="460" width="640">https://www.youtube.com/watch?v=16g6fEup2pQ</html5media>