DREAM

Danish Regenerative Engineering Alliance for Medicine

Recent News:

Novel SpCas9 variant with better gene editing efficiency created by the DREAM team (Feb. 2017)

The DREAM team have created a new SpCas9 variant with enhanced gene editing efficiency. This Cas9 variant was created by postdoc fellow Lin Lin from the DREAM team. To enhance gene editing efficiency, the SpCas9 protein was fused to E.Coli Recombinant protein A (RecA), an enzyme involved in DNA damage repair pathway in materials. Our group has shown that, fusing RecA to the SpCas9 can fine-tune the double-strand DNA damage repair pathway in mammalian cells towards non-homologous end joining. This system is useful for gene editing applications such gene knockout or deletion. The study has been accepted by the Journal of Biotechnology. This work was associated to the supports from the Lundbeck Foundation, the Danish Research Council and the Innovation Fund Denmark (BrainStem). http://www.sciencedirect.com/science/article/pii/S0168165617300871

 

Modelling and rescuing human FTD3 pathology by stem cell technology and CRISPR genome editing (Feb. 2017)

The BrainStem and DREAM research groups describe a patient iPSC-derived neuronal model for FTD3. This cellular model shows endosome abnormalities previously reported in patients. Furthermore, it provides insights into the role of impaired mitochondria function and imbalanced iron homeostasis in FTD3 pathology. All observed phenotypes were rescued in CRISPR/Cas9-edited isogenic controls.

Patient iPSC-Derived Neurons for Disease Modeling of Frontotemporal Dementia with Mutation in CHMP2B

Yu Zhang, Benjamin Schmid, Nanett K. Nikolaisen, Mikkel A. Rasmussen, Blanca I. Aldana, Mikkel Agger, Kirstine Calloe, Tina C. Stummann, Hjalte M. Larsen, Troels T. Nielsen, Jinrong Huang, Fengping Xu, Xin Liu, Lars Bolund, Morten Meyer, Lasse K. Bak, Helle S. Waagepetersen, Yonglun Luo, Jørgen E. Nielsen, The FReJA Consortium, Bjørn Holst, Christian Clausen, Poul Hyttel, Kristine K. Freude,

Stem Cell Reports. http://dx.doi.org/10.1016/j.stemcr.2017.01.012

 

 

Call for papers on "Genome Editing in Stem cells"

A special issue with focuses on genome editing in stem cells is now opened for paper submission at the journal "Stem Cell International". You can find more information about this issue through the following hyperlink: https://www.hindawi.com/journals/sci/si/402561/cfp/

Authors can submit their manuscripts through the Manuscript Tracking System at mts.hindawi.com/submit/journals/sci/gesc/.

Manuscript Due Friday, 23 June 2017 First Round of Reviews Friday, 15 September 2017 Publication Date Friday, 10 November 2017

Lead Guest Editor Yonglun Luo, Aarhus University, Aarhus, Denmark

Guest Editors Laurent Roybon, Lund University, Lund, Sweden Kristine Freude, Copenhagen University, Copenhagen, Denmark Guangqian Zhou, Shenzhen University, Shenzhen, China

 

 

New CRISPR/Cas9 sgRNA validation and selection system developed by the DREAM team

(Jan 14, 2016) DURING the last three years, the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein 9 system (CRISPR/Cas9) has rapidly become the most promising and powerful genome editing tool. With great efforts and contribution from numerous scientists around the world, the CRISPR/Cas9 system has now been developed and tested in almost all scenarios of genome and epigenome editing applications, such loss-of-function by targeted gene disruption, gain-of-function by targeted gene insertion, targeted gene repression or activation, epigenome editing etc.

Supported by the Danish Research Council for Independent Research and the Lundbeck Foundation, the DREAM team, lead by Associate Professor Yonglun Luo, is endeavoring to establish, improve and apply the CRISPR/Cas9 system for targeted gene editing and activation for studying the reprogramming and differentiation of human induced pluripotent stem cells. Furthermore, the DREAM team is also developing methods that can targeted modified the DNA methylome, which give great potential for studying stem cell reprogramming, differentiation and cancer therapy.

C-CheckOne challenge in the application of the CRISPR/Cas9 system is the validation and selection of functionally active small guide RNAs (sgRNA), which can efficiently direct the Cas9 or catalytically inactive Cas9 (dCas9) to the correct genomic loci. This week, the DREAM team published their dual-fluorescent system, named C-Check, in the scientific journal Cellular and Molecular Life Sciences [1]. The C-Check system contains two truncated and inactive green fluorescent genes (GFP), which contains homology sequences of 500 base pairs. Between the two truncated GFP genes, the C-Check vector contains a Golden-gate cloning site, where any target site can be inserted. Upon CRISPR/Cas9-mediated cleavage in the target site in the C-Check vector, the truncated GFP genes can be repaired by the single-strand annealing mechanism in the cells, which leads to the expression of a functional GFP protein (Illustrative Figure). With this system, the group demonstrated that sgRNAs, which are validated by the C-Check vectors, are also functional active at the endogenous genomic loci.

 

For more information about the C-Check vector, please read the related publication:

[1] Zhou et al. 2016. Enhanced genome editing in mammalian cells with a modified dual-fluorescent surrogate system. Cell Mol Life Sci. 2016 Jan 11. [Epub ahead of print]. PMID: 26755436

The DREAM team is partner of the BrainStem – Stem Cell Center of Excellence in Neurology

Associate Professor Yonglun Luo's group is now partner of the BrainStem – Stem Cell Center of Excellence in Neurology. The BrainStem is supported by the Innovation Fund Denmark, which will do research in neurodegenerative diseases over a 6 year period - from 2015 until 2020.

13thTransgenic Technology (TT2016) meeting

Associate Professor Yonglun Luo (Alun) is the invited speaker at the 13thTransgenic Technology (TT2016) meeting in Prague, Czech Republic, from March 20th-23rd, 2016.

Publications from Yonglun Luo's group

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PhD course "Genome engineering" is now opened

244-05/01 Genome engineering

Teach PhD students with the state-of-the-art knowledge on genome engineering using precision DNA nucleases, with a special focus on the RNA-guided DNA endonuclease CRISPR/Cas9. Systematically introduce what applications and scientific questions can be achieved and addresses by genome engineering, and thus inspire PhD students on their projects. Instruct PhD students on how design, generate, deliver and analyze CRISPR/Cas9 vector and activity. Provide valuable trouble shooting solutions on genome engineering. 

Course dates
06 March 2017 - 09 March 2017
Lecturer
Yonglun Luo
Place/Venue
Aarhus University 
City
Aarhus
ECTS
1,9 points

Link
service.health.au.dk/modules/Course/mypage/coursecalendar