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app note: comPOUND & arktic storage and cold sorting of clinical research samples in the laboratory for maximum sample integrity
blog: the perfect couple for GPCR.
Obtaining detailed structures of any membrane protein has been challenging. A detergent is required to extract the protein from the lipid environment which does not contribute to a favourable environment for crystallisation using traditional vapour diffusion methods. To overcome this problem the lipid cubic phase (LCP) crystallisation method was developed. LCP was originally limited only by the difficulties associated with handling and pipetting the extremely viscous cubic phase (monoolein and water) a hurdle that has now been overcome by the development of TTP Labtech’s robotic system, mosquito® LCP.
Crystallography has clearly made many contributions to science over the last century. The ongoing process of developing new therapeutic drugs has driven research towards a desire to understand drug-target interactions at a molecular level. One group of potential targets attracting a great deal of attention is the G-protein coupled receptors (GPCRs). GPCRs are a large and diverse group of eukaryotic membrane receptors that play a role in a plethora of biological functions from olfaction to mood regulation.
Many drugs exert their effects by binding to GPCRs and thus understanding the structure of this interaction is of great importance.
The use of mosquito LCP has alleviated many of the technical limitations around the crystallisation of membrane proteins and allowed the crystallography research community to successfully determine many new GPCR structures that relate to drug therapy. Below are a few examples of the these which have all used mosquito LCP as an essential tool in determining the structures.
anxiety and depression
Research at Heptares Therapeutics has resulted in several publications in Nature describing the crystal structures of class B and class C GPCRs. Corticotropin-releasing factor receptor type 1 (CRF1R) is a class B GPCR involved in mediating the body’s response to stress and has thus been a target of drugs designed to treat both anxiety and depression. This was the first crystal structure of a class B GPCR to be determined1. More recent work from Heptares has determined the crystal structure of a class C GPCR, metabotropic glutamate receptor 5 transmembrane domain (mGlu5). mGlu5 responds to the neurotransmitter glutamate, which may open up research into the treatment of fragile X syndrome, autism, depression, anxiety, addiction and movement disorders2.
type 2 diabetes
Type 2 diabetes affects over 3 million people in the UK alone and is the result of an insufficient insulin generation leading to higher than normal levels of blood glucose. A specific GPCR known as the human GPR40 receptor (hGPR40) has been shown to enhance glucose-dependent insulin secretion. Recent work has led to the development of TAK-875, a selective agonist of hGPR40 from Takeda that reached Phase III trials for the treatment of type 2 diabtes3. Crystallographic methods, involving the use of automated robotic systems (including mosquito), successfully described novel hGPR40-TAK-875 interactions4.
tackling blood clots
Recent work in crystallography has looked at a particular type of GPCR known as the P2Y12 receptor (P2Y12R), which is part of a family making up one of the most common targets of drugs that work to inhibit platelet aggregation5. Platelets play a critical role in thrombus formation (blood clotting), and drugs targeting P2Y12R have been approved for the prevention of stroke and myocardial infarction. Studies by Zhang et al revealed exciting and surprising structural findings:
P2Y12R was found to be capable of undergoing striking extracellular conformational changes akin to an open or closed ‘binding pocket’ depending on the type of agonist of antagonist present.
the integration of automation
Automated pipettors such as the mosquito range (TTP Labtech), used in all of the research detailed here, have been described as ‘the iPads of robotics’ by a recent user. Technology has allowed many users to progress from manually setting up crystallography screens and conducting multiple experiments in 24-well plates to rapidly seeding a 96-well screen. In the process the added advantages are the use of very low volumes of reagents with varying viscosities and, with a high degree of both accuracy and precision. These features have proven essential in taking the challenge of crystallising membrane proteins to a relatively routine procedure.
mastering the black art
Using these new systems has meant that even proteins that have been notoriously difficult to crystallise due to their complex structure, poor availability or unique characteristics, can now be studied. Whether using hanging drop, microbatch, vapour diffusion, sitting drop or lipid cubic phase (LCP) set-ups, there are now systems available to help alleviate many of the experimental hurdles experienced in the past.
For the full article please go to http://www.labnews.co.uk/features/the-perfect-couple/. If you would like any further information about any of the liquid handling mosquito or dragonfly® systems please contact firstname.lastname@example.org.
- Hollenstein, K. et al. Structure of class B GPCR corticotropin-releasing factor receptor 1. Nature 499, 438–43 (2013).
- Doré, A. S. et al. Structure of class C GPCR metabotropic glutamate receptor 5 transmembrane domain. Nature (2014). doi:10.1038/nature13396
- Negoro, N. et al. Discovery of TAK-875: A potent, selective, and orally bioavailable GPR40 agonist. ACS Med. Chem. Lett. 1, 290–294 (2010).
- Srivastava, A. et al. High-resolution structure of the human GPR40 receptor bound to allosteric agonist TAK-875. Nature (2014). doi:10.1038/nature13494
- Zhang, J. et al. Agonist-bound structure of the human P2Y12 receptor. Nature 509, 119–22 (2014).
STEM Visit to Alconbury Church of England Primary School for Science Week…
Our STEM ambassadors were invited via the Eastern Region STEM Networking Group to visit Alconbury Church of England Primary School in support of Science Week. Engineer Simon Tullett and scientist Dr Gillian Lewis attended from the TTP Labtech STEM Ambassador group to undertake some practical and fun science experiments and to demonstrate a liquid handling robot to the children.
Alconbury primary school is in a small village near to Huntingdon in Cambridgeshire and is a single form entry school catering for children from Reception through to Year 6 (4.5 to 11 years of age). Each class is approximately 30 children. On the day, we had the pleasure of 20, Year 6 children both boys and girls.
The day started with introductions about what STEM was and finding out what the children thought about the STEM subjects. We then quickly moved into our first activity of making bath bombs. Gillian explained that it was a demonstration of a chemical reaction that would be catalysed by the bath water. The children mixed citric acid and bicarbonate of soda with essential oils and food colouring to make a variety of sweet smelling and colourful bath bombs. The work involved carefully measuring out the ingredients and mixing properly before moulding them. The children and teachers all made bath bombs to take home that night.
Making bath bombs
The second activity was making and using invisible ink. This was made by mixing a base, in this case bicarbonate of soda with water. The children then wrote secret messages to each other (along the lines of “I hate my sister/brother”!) and allowed the ink to dry. To reveal the message an acid was used, in this case grapefruit juice to neutralise the base.
We then had a talk about the dragonfly robot and discussed what sort of skills people need to make a machine like this. We looked at the costs and timescales and explained that it isn’t just STEM subjects but a whole variety of other skills are needed when designing a machine. This was followed by a demonstration and each child got to drive the dragonfly and produce a brightly coloured dye pattern on a paper towel by running a gradient protocol on the dragonfly. The chemicals used in this experiment were bright inks from highlighter pens and this demonstrated how different quantities of material can very accurately be mixed together.
The final experiment of the day was to grow some crystals using a saturated solution of sugar. Each child made up a test tube with a wick dipped in the solution. The sugar crystals should start to form over the rest of the week for them.
We wrapped up with a Q&A session where the children asked lots of questions and told us about their experiences. One boy told us how his brother had undergone chemotherapy and was interested to learn how machines like dragonfly could be used in the fight to find a cure for cancer.
Simon and Gillian were invited to dinner on the promise that we come back and do something else at the school in the future!
ISBER 2017! Our travel grant winner, Brad Godfrey shares his experience of the event…
This year’s ISBER 2017 Annual Meeting took place in the beautiful city of Toronto, Canada. As well as an amazing location, the chance to network with peers and exciting talks, delegates were treated to a sell-out gala evening held at the spectacular Casa Loma – a romantic homage to the medieval era and one of Toronto’s top tourist attractions.
There were also a variety of awards to be had, most notably the award for ‘Outstanding Achievement in Biobanking’ Dr. Allison Hubel is Professor of Mechanical Engineering and Director of the Biopreservation Core Resource (BioCoR), a national resource in biopreservation. Her research focuses on development of fit-for-purpose protocols for preservation, development of technology to improve preservation/processing of cells, and understanding molecular mechanisms of damage during preservation.
A few brave souls also joined the ISBER 5K fun run in near darkness at dawn. The route wound through the local city sights and ended with winner Kaj Rydman being presented with a most unusual trophy…
One lucky delegate sampling ISBER’s delights was our travel grant winner Brad Godfrey from Michigan Medicine at the University of Michigan, USA who kindly shared his experience of the event:
Brad, what did you enjoy most about ISBER 2017?
Toronto was a wonderful city. I found the talk from the astrobiologist to be the most interesting; not necessarily the most useful, but definitely the most interesting and thought provoking!
So what are the latest trends that other biobanking specialists and researchers should keep an eye out for?
Everything is now automated. Biobankers should be able to program automation. In fact, I wish there were more vendors with automation at the conference and less with freezers.
Apart from some new regulatory changes, what else did you find from the conference that will be most relevant and/or useful to your research?
There were some wonderful ideas on quality control measures over time. And the networking was beneficial too.
What did you find most surprising about the event?
The amount of people that were not actually biobanking. There was a surprising amount of just regular bench scientists. Their collections were large for bench scientists but small for a biobank. And, if you don’t advertise/share your samples, it’s not really a biobank.
What (if anything) did you think was missing from the event?
I would like to have seen a talk on the effectiveness of different LIMS systems for different institutions. Some might be better at expanding collections / adding cohorts and others seem to be good at allowing programming for customization. An unbiased approach may have been difficult.
Could ISBER have been done anything better?
I was pretty happy with the overall experience.
Would it be worth going to the ISBER conference again next year?
I think not. Maybe every other year would be more beneficial.
Agreed, it could be perceived as expensive to go to every year, however for those unable to, this year ISBER held its first ever live webcast – streaming “Polar Shift: How Biobanking is Changing Our Thinking and the World”. Speakers included the keynote speaker Hannes Dempewolf (CropTrust, Bonn, Germany).
The talks highlighted how evidence derived from biobanking, and its associated practices, is shown to be instrumental in new discoveries, new processes, and new ways of thinking that are changing practices in medicine, agriculture and veterinary/environmental arenas. Being deemed a success, the live webcast may feature in future meetings and allow cash restricted labs to still engage with ISBER to gain something from the conference.
10 - 13 Sept 2017 Have you met dragonfly discovery? Join us at MipTec 2017
Come and see the team at MipTec 2017, booth #A26
Venue: Congress Center Basel, Switzerland
Date: 10 – 13 September 2017
Join us at this years MipTec and hear about the game-changing solution to challenges in assay development and high throughput screening – meet dragonfly discovery… any liquid, any volume, any well, any time!
dragonfly discovery gives scientists a common platform whereby they can easily develop complex assays, validate and screen them in a reliable, robust and cost efficient manner. Watch the video and discover dragonfly discovery’s innovative technology!
Watch the SelectScience interview with our Director, Product Strategy, Joby Jenkins about dragonfly discovery!
We will also be showcasing mirrorball, our high sensitivity fluorescence cytometer!
mirrorball is a is a highly versatile instrument that brings process efficiencies to multiple stages in the biologics screening pipeline – from early screening and hit identification, through to characterisation and phenotypic cellular responses.
Find out more about the mirrorball technology here!
Read the following poster:
- simultaneous determination of antibody binding, specificity and titre on the mirrorball fluorescence cytometer
Request the related app notes here:
- mirrorball: enhanced ELISA workflows with no-wash multiplexed fluorescent bead assays
- mirrorball: improving the reliability of cell-based immunoassay screening through multiplexing with mirrorball
Check in again soon for more information on what we will be showcasing and more!
For more information about MipTec click here
21 - 28 Aug 2017 Interested in mosquito crystal? Come and see us at IUCr 2017
Meet the team at IUCr 2017!
Venue: Hyderabad International Convention Centre, Hyderabad, India
Date: 21 – 28 August 2017
Join us at the 24th Congress & General Assembly of the International Union of Crystallography 2017 from 21 – 28 August 2017!
With mosquito crystal, you can use smaller volumes of precious protein sample with no risk of cross contamination. This results in cost savings and allows more extensive screening. You can automate all the popular protein crystallisation screening techniques – hanging drop, sitting drop and microbatch as well as seeding or additive screening plate preparation – without the need to make set-up changes to the instrument.
Read the app notes:
See our latest mosquito crystal related posters here:
Listen to great crystallography stories in 30 minutes: discover TTP Labtech’s webinar series
For more information about the event click here
Congratulations to the winner of our IUCr 2017 travel grant: Nicole Bertoletti
Hyderabad is a modern city of 9 million people in central India. Its iconic architectural symbol, the Charminar or ‘four minarets’ draws its
inspiration from the tetragonal crystal system. And fittingly, it’s against this exotic background that the 24th Congress & General Assembly of the International Union of Crystallography 2017 will be held this year. To support such an exciting event, we recently ran an IUCr 2017 travel grant competition and are excited to be able to share our interview with winner Nicole Bertoletti from Philipps-Universität Marburg in Germany:
Nicole, please tell us about your role and your research:
“I am a PhD student. I am about to finalize my dissertation in the research group of Prof. Dr. Klebe, in the department of Pharmaceutical Chemistry of the Philipps-University Marburg. During the course of my PhD, I obtained extensive theoretical and practical experience with various techniques including, but not limited to, recombinant protein expression and purification, bioinformatics as well as X-ray crystallography. The results of my work in these areas led up to now to two peer-reviewed publications in the Journal of Medicinal Chemistry. I worked on a new challenging project with the most recently identified member of the 17β-HSD protein family: the human 17β-HSD14. This protein has so far not been thoroughly investigated in details and its physiological role remains unknown. However, experiments revealed that h17β-HSD14 is predominantly expressed in brain and due to its turnover of estradiol it may become a potential target for the treatment of neuronal diseases for which estrogens were demonstrated to have neuroprotective effects. During the last three years, I established a successful expression and purification protocol for the recombinant h17β-HSD14 protein and I elucidated the first eight crystal structures of the protein in ternary complex with its cofactor and highly potent nonsteroidal inhibitors or estrone, the product of the catalysis reaction. Crystal structures of the protein-ligand complexes were necessary for the understanding of the inhibitors’ SAR (structure-activity relationship) and their further optimization. In fact, my studies revealed how small changes at the inhibitors’ substituents affect variations of their binding modes. In parallel, with the goal to discover new inhibitor scaffolds, we initiated a fragment-based lead discovery campaign by X-ray crystallography screening of a 96 fragment library.”
How does your research apply to the topics of discussion at the IUCr 2017 conference?
“I will actively contribute to the congress in the session M-009 “Enzymes, mechanism and drug design” with a poster presentation entitled: “X-ray Crystallography: essential tool for protein characterization and ligand optimization”. I would like to share with the crystallography community the importance of X-ray crystallography in medicinal chemistry for a rational design/optimization of ligands and its crucial role in the characterization of my target protein h17β-HSD14. We also believe that X-ray crystallographic fragment screening is a promising approach which may lead to the identification of more hits than other biophysical screening methods, especially for those ligands that have weak binding affinity, while providing essential structural information about binding modes necessary for their further optimization. In addition, my participation to the conference would allow me to discuss our findings, to present the binding mode of the crystallographic hits from our fragment screen against h17β-HSD14 and to discuss how I could use this information for the improvement of our inhibitors”
What do you hope to learn/gain from attending the conference?
“In the past years, I have already expressed my interest in the field of crystallography with my active participation of several national conferences and workshops, where I was even awarded with the MX-poster prize at the Seventh Joint BER II and BESSY II User Meeting 2015 in Berlin. The IUCr 24 meeting is the most important conference in the crystallography field with an expected attendance of over 2000 scientist from the entire world. With my scientific background, I believe that the participation at the IUCr 24 in Hyderabad would be an extraordinary chance for establish scientific connection with experts in the field with whom I would like to share the results, discuss challenges and the experience that I acquired during my work. The IUCr 24 congress would be a superb opportunity where I could have the possibility to broaden my view on in the macromolecular crystallography field and to keep track of new developments. In fact, the conference offers an impressive scientific program that covers many areas and aspects of macromolecular crystallography and will also include presentation from industrial researchers. The participation of the IUCr meeting would not only be a great opportunity for me, but also for the entire lab that could benefit from the new knowledge that I will acquire and communicate once back home. In addition, I aspire to pursue an academic career in this field and I believe that the participation in scientific congress is crucial for this pursuit.”
Again our congratulations go out to Nicole and we look forward to welcoming both her and other crystallographers to the IUCr 2017 conference in Hyderabad where we can be found at booth number #59 & 61.
ISBER 2017 travel grant winner: Brad Godfrey
As a global biobanking organisation, ISBER creates opportunities for networking, education, and brings together innovative approaches to evolving challenges in biological and environmental repositories. However, it’s also known as the ‘the place to be’ in the biobanking calendar and this year’s conference theme is ‘Due North: Aligning Biobanking Practice with Evolving Evidence and Innovation’. To support such an exciting event, we recently ran an ISBER 2017 travel grant competition and are excited to be able to share our interview with winner Brad Godfrey from Michigan Medicine at the University of Michigan, USA.
Brad, please tell us about your role and your research:
“I am the Lab Manager for several chronic kidney disease studies at the Applied Systems Biology Core at Michigan Medicine. I receive clinical biospecimen shipments from various sites around the world and I will catalog and store them. Part of my role is processing the tissues to ultimately perform transciptomics profiling on the compartments of the kidney and whole genome sequencing on the blood. I also provide blood and urine samples to our ancillary studies to perform other types of molecular characterization.”
“I work entirely with human patient samples. We take a complete -omics approach to kidney disease starting with protocol biopsies and correlating tissue transcriptomics data with longitudinal samples or urine and blood. We have a collection of over 250,000 urine and blood samples across several chronic kidney disease cohorts, collected from patients for over a decade, that we use ourselves or ship to ancillary studies throughout the world. Our interdisciplinary research team integrates information from a wide spectrum of human cohort studies we have initiated or are intimately involved with. In these prospective cohort studies, we test the precision medicine concept for renal disease by integrating information along the genotype-phenotype continuum using carefully monitored environmental exposures, genetic predispositions, epigenetic markers, transcriptional networks, proteomic profiles, metabolic fingerprints, digital histological biopsy archive, and prospective clinical disease characterization.”
What do you hope to learn/gain from attending the conference?
“I have several goals for attending ISBER. First, learn any regulatory changes that have recently occurred. We deal with human tissue exclusively and need to be aware of the latest regulations. I will get updated on the latest trends in biobanking and what our outside studies will expect when requesting samples. Knowing the most effective approaches will help further the biobanking effort. We collect various biosamples from low and moderate areas of the world. I will be better able to communicate with our clinical coordinators how to properly obtained, preserve, store, and ship these samples under less-than-ideal circumstances. This is crucial in our goals to study kidney disease such as HIV-induced nephropathy that disproportionately affects less advantaged areas.”
Again our congratulations go out to Brad and we look forward to hearing about his experience and key findings from the ISBER 2017 conference!
Going to ISBER 2017?
Come and meet our team at booth #30!
Find out more here
press release: Advanced Analytical Technologies and TTP Labtech Alliance Delivers High-Throughput NGS Library Prep Solutions
press release: Advanced Analytical Technologies and TTP Labtech Alliance Delivers High-Throughput NGS Library Prep Solutions
- The partnership combines mosquito® and Fragment Analyzer™ platforms to reduce cost, maximise throughput and improve quality of NGS library preparation.
- Stanford University teams published the novel high-throughput library prep workflows for single-cell RNA sequencing (scRNA-Seq) in Cell1 and Nature Scientific Data2.
Cambridge, UK, and Ankeny, IA, USA, 29th March 2017: TTP Labtech Ltd. is a global leader in the design and development of automated instrumentation and consumables for life science applications. Advanced Analytical Technologies, Inc. (AATI) is the award-winning manufacturer of Fragment Analyzer™ and FEMTO Pulse™ automated systems for nucleic acid sizing and quantity analysis. The Fragment Analyzer INFINITY™ model integrates within robotic cells for complete 24+ hour automation. The companies have signed a co-marketing agreement to support joint solutions that miniaturise and optimise high-throughput next-generation sequencing (NGS) library preparation workflows.
The new NGS methods, published by Stanford researchers in Cell (Loh and Chen, et al., 2016) and Nature Scientific Data (Koh and Sinha, et al., 2016), combine the versatile
TTP Labtech mosquito® automated low-volume liquid handlers with Advanced Analytical’s Fragment Analyzer™. The approach overcomes challenges associated with high-throughput single-cell sequencing. Combined use of the mosquito and Fragment Analyzer systems enables scale with cost-efficiency, accuracy, and precision.
Rahul Sinha, PhD, Prof. Irving Weissman Lab, Stanford University, said: “This semi-automated, robust SMART-seq2 workflow for single-cell RNA-seq has reduced our hands-on time from two weeks to a couple of days, while increasing the accuracy and lowering the cost. The AATI Fragment Analyzer and TTP Labtech’s mosquito X1 and mosquito HTS have been essential for this new workflow.”
The Stanford team’s mapping of stem cell differentiation along multiple mesodermal lineages required preparation of miniaturised RNA-Seq libraries from nearly a thousand single-cells, a demanding task that would have been difficult to achieve without high-throughput instrumentation and nanolitre-scale liquid handling. TTP Labtech’s mosquito liquid handlers leverage precise and accurate true-positive displacement technology. The mosquito X1 and HTS were used to prepare input plates for the Fragment Analyzer, automate cDNA normalisation and generate low-volume Nextera® XT sequencing libraries in 384-well format. Efficient and accurate quantification and quality analysis on the AATI Fragment Analyzer was essential for the large-scale scRNA-Seq application.
Jonathan Hagopian, PhD, Director of Business Development at Advanced Analytical, said: “This partnership with TTP Labtech is enabling high-throughput NGS discovery with robust miniaturisation and accurate quantification on the Fragment Analyzer. The low-volume liquid handling capabilities of the mosquito also empower novel applications with ultra-sensitive analysis on our new FEMTO Pulse system.”
Klaus Hentrich, Genomics Product Manager at TTP Labtech, added: “Precise nanolitre scale liquid handling combined with accurate high-sensitivity quantification is key for miniaturising NGS library prep. TTP Labtech and Advanced Analytical instruments, together, provide powerful cost-effective genomics workflow solutions.”
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Tel: +44 (0)7811 996 942
About TTP Labtech
TTP Labtech designs and manufactures robust, reliable and easy-to-use solutions for sample management, liquid handling and multiplexed detection in drug discovery and genomics. We enable life scientists, through collaboration, deep application knowledge and leading engineering, to accelerate research and make a difference together. Our essential tools include state-of-the-art solutions developed for high throughput compound and biologics screening (acumen® Cellista, mirrorball® and fully validated consumables such as sol-R™ beads and plates),flexible sample management workflows from ambient to -800C (comPOUND®, arktic®, lab2lab), and unique low-volume liquid handling for genomics, compound screening and protein crystallography (mosquito® X1, mosquito® HTS, mosquito® Crystal, mosquito® LCP, mosquito® HV, dragonfly® crystal and dragonfly® discovery and a full range of validated consumables such as tips and plates).
About Advanced Analytical Technologies https://www.aati-us.com/
Advanced Analytical Technologies, Inc. (AATI) develops, manufactures, and markets low and high-throughput automated analysis systems. The company’s instrument platforms optimize and accelerate complex workflows for basic science and commercial applications in life-science industries including genomics, molecular diagnostics, pharmaceuticals, healthcare, biotechnology, synthetic biology, biofuels, and agriculture. AATI’s product portfolio has instruments for parallel analysis of DNA, RNA, pharmaceutical compounds, proteins, and post-translational modifications using capillary electrophoresis (CE) with fluorescence and UV detection. The Fragment Analyzer™, Fragment Analyzer INFINITY™, and FEMTO Pulse™ are automated systems for sizing and concentration analysis of various DNA and RNA samples including genomic DNA, NGS libraries, CRISPR mutations, dsDNA fragments, PCR amplicons, microsatellite SSR, RFLP, total RNA, mRNA, small and microRNA, single-cell and cell-free isolates. Advanced Analytical has facilities in Ankeny, Iowa, USA, Heidelberg, Germany, and Paris, France.
- Kyle M. Loh*, Angela Chen*, Pang Wei Koh, Tianda Z. Deng, Rahul Sinha, Jonathan M. Tsai, Amira A. Barkal, Kimberle Y. Shen, Rajan Jain, Rachel M. Morganti, Ng Shyh-Chang, Nathaniel B. Fernhoff, Benson M. George, Gerlinde Wernig, Rachel E.A. Salomon, Zhenghao Chen, Hannes Vogel, Jonathan A. Epstein, Anshul Kundaje, William S. Talbot, Philip A. Beachy, Lay Teng Ang^, Irving L. Weissman^; Mapping the pairwise choices leading from pluripotency to human bone, heart, and other mesoderm cell types. Cell, 166:451-467, 2016.
- Pang Wei Koh*, Rahul Sinha*, Amira A. Barkal, Rachel M. Morganti, Angela Chen, Irving L. Weissman^, Lay Teng Ang^, Anshul Kundaje^ & Kyle M. Loh^; An atlas of transcriptional, chromatin accessibility, and surface marker changes in human mesoderm development. Nature Scientific Data, 3:160109, 2016.