Genomic Diagnostics: Three Techniques to Perform Single Cell Gene Expression and Genome Sequencing Single Molecule DNA Sequencing
Curator: Aviva Lev-Ari, PhD, RN
4.2.3 Genomic Diagnostics: Three Techniques to Perform Single Cell Gene Expression and Genome Sequencing Single Molecule DNA Sequencing, Volume 2 (Volume Two: Latest in Genomics Methodologies for Therapeutics: Gene Editing, NGS and BioInformatics, Simulations and the Genome Ontology), Part 4: Single Cell Genomics
This article presents Three Techniques to Perform Single Cell Gene Expression and Genome Sequencing Single molecule DNA sequencing
-
Stephen Quake, Quake Lab @Stanford University
-
Ido Amit, AmitLab @Department of Immunology at the Weizmann Institute of Science, Rehovot, Israel
-
Gilad Evrony – EPPENDORF 2016 GRAND PRIZE WINNER
Quake Lab @Stanford University
- Immune repertoire sequencing
- Single-cell sequencing
- Cell-free nucleic acid sequencing
Quake Lab Publications
Ido Amit, AmitLab @Department of Immunology at the Weizmann Institute of Science, Rehovot, Israel
The Amit lab studies the genomic code enabling immune cells to differentiate to specific subtypes and devise a specific response to invading pathogens. Our main focus is to understand how gene regulatory networks activate this code. We develop and apply state of the art high throughputgenomic tools and interdisciplinary approaches to address these critical biological and therapeutic questions. We believe that elucidating the underlying principles of the regulatory code will allow us to impact the future of personalized medicine. We are currently seeking highly motivated individuals who want to join us on this quest.
SOURCE
https://www.weizmann.ac.il/immunology/AmitLab/front
Immunology, one cell at a time
03 July 2017
Basic lessons – “Single-cell genomics will soon be commonplace in basic and applied immunology research.”
It is early days for single-cell genomics. But already, a number of important lessons can be learnt from the experiences of our lab and those of others.
First, it is clear that many of the current categories of immune cells, such as T cells or monocytes, encompass heterogeneous populations. To probe cellular complexity, researchers must therefore cast their nets wide, and try to collect all immune cells within a tissue or region of interest. This is a very different approach from that used with methods based on cell-surface markers, which aim to obtain as pure a sample as possible.
Second, success will depend, in part, on the extent to which researchers preserve the states of cells and the original composition of a tissue. Cell stress or death should be minimized to ensure that tissue preparation does not favour specific cell types. (Some are more sensitive to heat stress, for example, than others.)
Third, bioinformaticians will need to develop scalable and robust algorithms to cope with greater numbers of cells, conflicting or overlapping programs of gene expression and fleeting developmental stages.
Fourth, after researchers have characterized all of the immune cells in a sample, they will need to find molecular markers that can be used to either enrich or deplete certain cell types in further samples. Tissues comprise trillions of cells with myriad molecular characteristics and functions, and the types or states of these cells may vary in abundance by many orders of magnitude. For instance, in the brains of healthy mice, our newly identified population of DAM makes up less than 0.01% of cells15. Thus, repeated unbiased sampling to characterize rare populations will keep on accumulating cells that are not those of interest.
Nature 547, 27–29 (06 July 2017) doi:10.1038/547027a
http://www.nature.com/news/immunology-one-cell-at-a-time-1.22232?WT.ec_id=NEWSDAILY-20170703
Ido Amit – Selected References
Innate Immune Landscape in Early Lung Adenocarcinoma by Paired Single-Cell Analyses.
Lavin Y, Kobayashi S, Leader A, Amir ED, Elefant N, Bigenwald C, Remark R, Sweeney R, Becker CD, Levine JH, Meinhof K, Chow A, Kim-Shulze S, Wolf A, Medaglia C, Li H, Rytlewski JA, Emerson RO, Solovyov A, Greenbaum BD, Sanders C, Vignali M, Beasley MB, Flores R, Gnjatic S, Pe’er D, Rahman A, Amit I, Merad M.
Cell. 2017 May 4;169(4):750-765.e17. doi: 10.1016/j.cell.2017.04.014.
- PMID:
- 28475900
MicroRNA-142 controls thymocyte proliferation.
Mildner A, Chapnik E, Varol D, Aychek T, Lampl N, Rivkin N, Bringmann A, Paul F, Boura-Halfon S, Hayoun YS, Barnett-Itzhaki Z, Amit I, Hornstein E, Jung S.
Eur J Immunol. 2017 May 4. doi: 10.1002/eji.201746987. [Epub ahead of print]
- PMID:
- 28471480
Single-cell transcriptome conservation in cryopreserved cells and tissues.
Guillaumet-Adkins A, Rodríguez-Esteban G, Mereu E, Mendez-Lago M, Jaitin DA, Villanueva A, Vidal A, Martinez-Marti A, Felip E, Vivancos A, Keren-Shaul H, Heath S, Gut M, Amit I, Gut I, Heyn H.
Genome Biol. 2017 Mar 1;18(1):45. doi: 10.1186/s13059-017-1171-9.
- PMID:
- 28249587
Single-cell spatial reconstruction reveals global division of labour in the mammalian liver.
Bahar Halpern K, Shenhav R, Matcovitch-Natan O, Tóth B, Lemze D, Golan M, Massasa EE, Baydatch S, Landen S, Moor AE, Brandis A, Giladi A, Stokar-Avihail A, David E, Amit I, Itzkovitz S.
Nature. 2017 Feb 16;542(7641):352-356. doi: 10.1038/nature21065. Epub 2017 Feb 6. Erratum in: Nature. 2017 Mar 30;543(7647):742.
- PMID:
- 28166538
Gilad Evrony, MD, PhD
EPPENDORF 2016 GRAND PRIZE WINNER
Gilad Evrony

+ See all authors and affiliations
Summary
We each begin life as a single cell harboring a single genome, which—over the course of development—gives rise to the trillions of cells that make up the body. From skin cells to heart cells to neurons of the brain, each bears a copy of the original cell’s genome. But as anyone who has used a copy machine or played the childhood game of “telephone” knows, copies are never perfect. Every cell in an individual actually has a unique genome, an imperfect copy of its cellular ancestor differentiated by inevitable somatic mutations arising from errors in DNA replication and other mutagenic forces (1). Somatic mutation is the fundamental process leading to all genetic diseases, including cancer; every inherited genetic disease also has its origins in such mutation events that occurred in an ancestor’s germline cells. Yet how many and what kinds of somatic mutations accumulate in our cells as we develop and age has long been unknown and a blind spot in our understanding of the origins of genetic disease.
(1)
http://science.sciencemag.org/content/sci/354/6312/557.full.pdf
The author of the prize-winning essay, Gilad Evrony, received his undergraduate degree from the Massachusetts Institute of Technology. He served in the Intelligence Division of the Israel Defense Forces and completed an M.D. and Ph.D. at Harvard Medical School, with graduate research in the laboratory of Dr. Christopher Walsh at Boston Children’s Hospital. Dr. Evrony is currently pursuing clinical training in pediatrics at Mount Sinai Hospital and continuing his research developing novel technologies for studying the brain and neuropsychiatric diseases.
https://www.sciencemag.org/prize/eppendorf/gilad-evrony
Single-neuron genome sequencing is revealing clues about what goes wrong in the brain
https://www.technologyreview.com/lists/innovators-under-35/2015/pioneer/gilad-evrony/
Gilad Evrony – PubMed Selected Publications
Evrony GD, Cordero DR, Shen J, Partlow JN, Yu TW, Rodin RE, Hill RS, Coulter ME, Lam AN, Jayaraman D, Gerrelli D, Diaz DG, Santos C, Morrison V, Galli A, Tschulena U, Wiemann S, Martel MJ, Spooner B, Ryu SC, Elhosary PC, Richardson JM, Tierney D, Robinson CA, Chibbar R, Diudea D, Folkerth R, Wiebe S, Barkovich AJ, Mochida GH, Irvine J, Lemire EG, Blakley P, Walsh CA.
Genome Res. 2017 Jun 19. doi: 10.1101/gr.219899.116. [Epub ahead of print]
- PMID:
- 28630177
Evrony GD.
Science. 2016 Nov 4;354(6312):557-558. No abstract available.
- PMID:
- 27811258
Resolving rates of mutation in the brain using single-neuron genomics.
Evrony GD, Lee E, Park PJ, Walsh CA.
Elife. 2016 Feb 22;5. pii: e12966. doi: 10.7554/eLife.12966.
- PMID:
- 26901440
Somatic mutation in single human neurons tracks developmental and transcriptional history.
Lodato MA, Woodworth MB, Lee S, Evrony GD, Mehta BK, Karger A, Lee S, Chittenden TW, D’Gama AM, Cai X, Luquette LJ, Lee E, Park PJ, Walsh CA.
Science. 2015 Oct 2;350(6256):94-8. doi: 10.1126/science.aab1785.
- PMID:
- 26430121
Cai X, Evrony GD, Lehmann HS, Elhosary PC, Mehta BK, Poduri A, Walsh CA.
Cell Rep. 2015 Feb 3;10(4):645. doi: 10.1016/j.celrep.2015.01.028. Epub 2015 Feb 3. No abstract available.
- PMID:
- 25832109