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Stem Cell Heterogeneity

Overview of attention for book
Cover of 'Stem Cell Heterogeneity'

Table of Contents

  1. Altmetric Badge
    Book Overview
  2. Altmetric Badge
    Chapter 315 Isolation and Culture of Embryonic Stem Cells, Mesenchymal Stem Cells, and Dendritic Cells from Humans and Mice
  3. Altmetric Badge
    Chapter 319 Maintenance of Dermal Papilla Cells by Wnt-10b In Vitro
  4. Altmetric Badge
    Chapter 320 Maintenance of Skin Epithelial Stem Cells by Wnt-3a In Vitro
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    Chapter 321 Isolation and Expansion of Muscle Precursor Cells from Human Skeletal Muscle Biopsies
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    Chapter 322 An Effective and Reliable Xeno-free Cryopreservation Protocol for Single Human Pluripotent Stem Cells
  7. Altmetric Badge
    Chapter 323 Induction of a Tumor-Metastasis-Receptive Microenvironment as an Unwanted Side Effect After Radio/Chemotherapy and In Vitro and In Vivo Assays to Study this Phenomenon
  8. Altmetric Badge
    Chapter 324 Decoding the Epigenetic Heterogeneity of Human Pluripotent Stem Cells with Seamless Gene Editing
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    Chapter 325 Stencil Micropatterning for Spatial Control of Human Pluripotent Stem Cell Fate Heterogeneity
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    Chapter 326 Isolation and Characterization of Cancer Stem Cells of the Non-Small-Cell Lung Cancer (A549) Cell Line
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    Chapter 327 Aerosol-Based Cell Therapy for Treatment of Lung Diseases
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    Chapter 327 Erratum to: Enzyme-Free Dissociation of Neurospheres by a Microfluidic Chip-Based Method
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    Chapter 328 Induction of Inner Ear Hair Cells from Mouse Embryonic Stem Cells In Vitro
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    Chapter 329 In Vitro Culture of Human Hematopoietic Stem Cells in Serum Free Medium and Their Monitoring by Flow Cytometry
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    Chapter 342 Isolation and Propagation of Glioma Stem Cells from Acutely Resected Tumors
  16. Altmetric Badge
    Chapter 343 Clonal Analysis of Cells with Cellular Barcoding: When Numbers and Sizes Matter
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    Chapter 344 Automated Cell-Based Quantitation of 8-OHdG Damage
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    Chapter 345 Heterogeneity of Stem Cells: A Brief Overview
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    Chapter 346 Agent-Based Modeling of Cancer Stem Cell Driven Solid Tumor Growth.
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    Chapter 347 Establishment and Characterization of Naïve Pluripotency in Human Embryonic Stem Cells.
  21. Altmetric Badge
    Chapter 348 Enzyme-Free Dissociation of Neurospheres by a Microfluidic Chip-Based Method
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    Chapter 349 Visualizing the Functional Heterogeneity of Muscle Stem Cells
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    Chapter 350 CoCl2 Administration to Vascular MSC Cultures as an In Vitro Hypoxic System to Study Stem Cell Survival and Angiogenesis
  24. Altmetric Badge
    Chapter 356 Dissecting Transcriptional Heterogeneity in Pluripotency: Single Cell Analysis of Mouse Embryonic Stem Cells
  25. Altmetric Badge
    Chapter 357 Generation of Regionally Specific Neural Progenitor Cells (NPCs) and Neurons from Human Pluripotent Stem Cells (hPSCs)
  26. Altmetric Badge
    Chapter 358 Measuring ATP Concentration in a Small Number of Murine Hematopoietic Stem Cells.
  27. Altmetric Badge
    Chapter 360 Reporter Systems to Study Cancer Stem Cells
  28. Altmetric Badge
    Chapter 361 Analysis of Cell Cycle Status of Murine Hematopoietic Stem Cells
  29. Altmetric Badge
    Chapter 362 Growth Factor-Free Pre-vascularization of Cell Sheets for Tissue Engineering
Attention for Chapter 324: Decoding the Epigenetic Heterogeneity of Human Pluripotent Stem Cells with Seamless Gene Editing
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About this Attention Score

  • In the top 25% of all research outputs scored by Altmetric
  • Good Attention Score compared to outputs of the same age (79th percentile)
  • High Attention Score compared to outputs of the same age and source (93rd percentile)

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Chapter title
Decoding the Epigenetic Heterogeneity of Human Pluripotent Stem Cells with Seamless Gene Editing
Chapter number 324
Book title
Stem Cell Heterogeneity
Published in
Methods in molecular biology, April 2016
DOI 10.1007/7651_2016_324
Pubmed ID
Book ISBNs
978-1-4939-6549-6, 978-1-4939-6550-2
Authors

Singh, Amar M, Perry, Dustin W, Steffey, Valeriya V Adjan, Miller, Kenneth, Allison, Daniel W, Amar M. Singh, Dustin W. Perry, Valeriya V. Adjan Steffey, Kenneth Miller, Daniel W. Allison, Singh, Amar M., Perry, Dustin W., Steffey, Valeriya V. Adjan, Allison, Daniel W.

Abstract

Pluripotent stem cells exhibit cell cycle-regulated heterogeneity for trimethylation of histone-3 on lysine-4 (H3K4me3) on developmental gene promoters containing bivalent epigenetic domains. The heterogeneity of H3K4me3 can be attributed to Cyclin-dependent kinase-2 (CDK2) phosphorylation and activation of the histone methyltransferase, MLL2 (KMT2B), during late-G1. The deposition of H3K4me3 on developmental promoters in late-G1 establishes a permissive chromatin architecture that enables signaling cues to promote differentiation from the G1 phase. These data suggest that the inhibition of MLL2 phosphorylation and activation will prevent the initiation of differentiation. Here, we describe a method to seamlessly modify a putative CDK2 phosphorylation site on MLL2 to restrict its phosphorylation and activation. Specifically, by utilizing dimeric CRISPR RNA-guided nucleases, RFNs (commercially known as the NextGEN™ CRISPR), in combination with an excision-only piggyBac™ transposase, we demonstrate how to generate a point mutation of threonine-542, a predicted site to prevent MLL2 activation. This gene editing method enables the use of both positive and negative selection, and allows for subsequent removal of the donor cassette without leaving behind any unwanted DNA sequences or modifications. This seamless "donor-excision" approach provides clear advantages over using single stranded oligo-deoxynucleotides (ssODN) as donors to create point mutations, as the use of ssODN necessitate additional mutations in the donor PAM sequence, along with extensive cloning efforts. The method described here therefore provides the highest targeting efficiency with the lowest "off-target" mutation rates possible, while removing the labor-intensive efforts associated with screening thousands of clones. In sum, this chapter describes how seamless gene editing may be utilized to examine stem cell heterogeneity of epigenetic marks, but is also widely applicable for performing precise genetic manipulations in numerous other cell types.

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X Demographics

The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 30 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United Kingdom 1 3%
Unknown 29 97%

Demographic breakdown

Readers by professional status Count As %
Researcher 9 30%
Student > Ph. D. Student 5 17%
Other 4 13%
Student > Bachelor 3 10%
Professor 2 7%
Other 2 7%
Unknown 5 17%
Readers by discipline Count As %
Agricultural and Biological Sciences 11 37%
Biochemistry, Genetics and Molecular Biology 8 27%
Medicine and Dentistry 2 7%
Mathematics 1 3%
Immunology and Microbiology 1 3%
Other 1 3%
Unknown 6 20%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 8. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 31 May 2016.
All research outputs
#3,983,974
of 22,862,742 outputs
Outputs from Methods in molecular biology
#1,032
of 13,127 outputs
Outputs of similar age
#62,703
of 300,620 outputs
Outputs of similar age from Methods in molecular biology
#2
of 29 outputs
Altmetric has tracked 22,862,742 research outputs across all sources so far. Compared to these this one has done well and is in the 82nd percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 13,127 research outputs from this source. They receive a mean Attention Score of 3.4. This one has done particularly well, scoring higher than 92% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 300,620 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 79% of its contemporaries.
We're also able to compare this research output to 29 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 93% of its contemporaries.