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 Thanks to multiple advantages, the CRISPR system superseded its predecessors and became a popular method for genome editing.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 ==
-=Cardiovascular and Placenta=
-[[File:Placental membranes.jpg|thumb|300px|alt=Fetal membranes and Placenta|Fetal membranes and Placenta]]
-== Introduction ==
-This laboratory will explore the development of the embryonic cardiovascular system, which includes the placental vascular system. Placenta development showing both fetal and maternal changes will be shown  followed by an overview of the flow of blood through the embryonic vasculature. Examples include histological sections of both placenta, uterus and the early embryo (Carnegie stage 13) cross-sections.
-==Objectives==
+== 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
+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
-# Understand the early placentation events and the development of placental blood vessels.
+-      Development of new assisted reproductive techniques
-# Brief understanding of prenatal diagnosis and cord stem cells.
-# Understand development of early embryo heart and vascular system.
-==References==
+-      Discovery and engineering of new endonucleases
-{|
+-      Investigating the pathomechanisms of FoxG1 syndrome
-| {{Embryo logocitation}}
-|
-{{Placenta Links}}
-{{Heart Links}}
-|-
-| {{MPT2011cover_citation}}
-| The following chapter links only work with a UNSW connection.
-* [http://www.unsw.eblib.com.wwwproxy0.library.unsw.edu.au/patron/Read.aspx?p=1430154&pg=131 Chapter 7 - Placenta and Fetal Membranes]
-* [http://www.unsw.eblib.com.wwwproxy0.library.unsw.edu.au/patron/Read.aspx?p=1430154&pg=311 Chapter 13 - Cardiovascular System]
-|-
-| {{SBBF2009cover_citation}}
-| The following chapter links only work with a UNSW [http://er.library.unsw.edu.au/er/cgi-bin/eraccess.cgi?url=http://www.unsw.eblib.com.wwwproxy0.library.unsw.edu.au/patron/FullRecord.aspx?p=2074524 UNSW Library subscription]
-* Chapter 2 - Second Week: Becoming Bilaminar and Fully Implanting
-* Chapter 12 - Development of the Heart
-* Chapter 13 - Development of the Vasculature
 |}
-===Additional Textbooks===
+'''Lecture slides:''' [[File:Delerue.pdf]]
-[[File:Gray0039.gif|thumb|300px|alt=Image section through the placenta|Section through the placenta]]
+===References===
-* '''Human Embryology'''  Larson Ch7 p151-188 Heart, Ch8 p189-228 Vasculature
-* The Developing Human: Clinically Oriented Embryology (6th ed.) Moore and Persaud Ch14: p304-349
-* Before we Are Born (5th ed.) Moore and Persaud Ch12; p241-254
-* Essentials of Human Embryology Larson Ch7 p97-122 Heart, Ch8 p123-146 Vasculature
-* Human Embryology Fitzgerald and Fitzgerald Ch13-17: p77-111
+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]
+:"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 [https://dx.doi.org/10.1007/s00401-015-1486-0]
-{{ANAT2341Lab4}}
+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]
+===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
+|}
+===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]
-{{2015ANAT2341 footer}}
+<html5media height="460" width="640">https://www.youtube.com/watch?v=16g6fEup2pQ</html5media>