Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains
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Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains. / Jurisic, Anamarija; Robin, Chloé; Tarlykov, Pavel; Siggens, Lee; Schoell, Brigitte; Jauch, Anna; Ekwall, Karl; Sørensen, Claus Storgaard; Lipinski, Marc; Shoaib, Muhammad; Ogryzko, Vasily.
I: Nucleic Acids Research, Bind 46, Nr. 22, e135, 2018, s. 1-16.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains
AU - Jurisic, Anamarija
AU - Robin, Chloé
AU - Tarlykov, Pavel
AU - Siggens, Lee
AU - Schoell, Brigitte
AU - Jauch, Anna
AU - Ekwall, Karl
AU - Sørensen, Claus Storgaard
AU - Lipinski, Marc
AU - Shoaib, Muhammad
AU - Ogryzko, Vasily
PY - 2018
Y1 - 2018
N2 - Analysis of large-scale interphase genome positioning with reference to a nuclear landmark has recently been studied using sequencing-based single cell approaches. However, these approaches are dependent upon technically challenging, time consuming and costly high throughput sequencing technologies, requiring specialized bioinformatics tools and expertise. Here, we propose a novel, affordable and robust microscopy-based single cell approach, termed Topokaryotyping, to analyze and reconstruct the interphase positioning of genomic loci relative to a given nuclear landmark, detectable as banding pattern on mitotic chromosomes. This is accomplished by proximity-dependent histone labeling, where biotin ligase BirA fused to nuclear envelope marker Emerin was coexpressed together with Biotin Acceptor Peptide (BAP)-histone fusion followed by (i) biotin labeling, (ii) generation of mitotic spreads, (iii) detection of the biotin label on mitotic chromosomes and (iv) their identification by karyotyping. Using Topokaryotyping, we identified both cooperativity and stochasticity in the positioning of emerin-associated chromatin domains in individual cells. Furthermore, the chromosome-banding pattern showed dynamic changes in emerin-associated domains upon physical and radiological stress. In summary, Topokaryotyping is a sensitive and reliable technique to quantitatively analyze spatial positioning of genomic regions interacting with a given nuclear landmark at the single cell level in various experimental conditions.
AB - Analysis of large-scale interphase genome positioning with reference to a nuclear landmark has recently been studied using sequencing-based single cell approaches. However, these approaches are dependent upon technically challenging, time consuming and costly high throughput sequencing technologies, requiring specialized bioinformatics tools and expertise. Here, we propose a novel, affordable and robust microscopy-based single cell approach, termed Topokaryotyping, to analyze and reconstruct the interphase positioning of genomic loci relative to a given nuclear landmark, detectable as banding pattern on mitotic chromosomes. This is accomplished by proximity-dependent histone labeling, where biotin ligase BirA fused to nuclear envelope marker Emerin was coexpressed together with Biotin Acceptor Peptide (BAP)-histone fusion followed by (i) biotin labeling, (ii) generation of mitotic spreads, (iii) detection of the biotin label on mitotic chromosomes and (iv) their identification by karyotyping. Using Topokaryotyping, we identified both cooperativity and stochasticity in the positioning of emerin-associated chromatin domains in individual cells. Furthermore, the chromosome-banding pattern showed dynamic changes in emerin-associated domains upon physical and radiological stress. In summary, Topokaryotyping is a sensitive and reliable technique to quantitatively analyze spatial positioning of genomic regions interacting with a given nuclear landmark at the single cell level in various experimental conditions.
U2 - 10.1093/nar/gky818
DO - 10.1093/nar/gky818
M3 - Journal article
C2 - 30215776
VL - 46
SP - 1
EP - 16
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 22
M1 - e135
ER -
ID: 203593193