Imaging of native transcription factors and histone phosphorylation at high resolution in live cells

Conic, Sascha; Desplancq, Dominique; Ferrand, Alexia; Fischer, Veronique; Heyer, Vincent; Reina‐San‐Martin, Bernardo; Pontabry, Julien; Oulad‐Abdelghani, Mustapha; N., Kishore Babu; Wright, Graham D.; Molina, Nacho; Weiss, Étienne; Tora, Làszlò

doi:10.1083/jcb.201709153

Cited by 36 publications

(39 citation statements)

References 69 publications

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“…The delivered antibody should show the same localization pattern as immunofluorescence of the target protein using a different antibody. It is also possible to fluorescently label the antibody in order to observe correct binding to target protein in live cells 27 . However, in some cases antibody delivery may cause target protein degradation via endogenous TRIM21 and/or cause target protein mislocalisation due to inhibitory effects of antibody binding.…”

Section: Experimental Designmentioning

confidence: 99%

Acute and rapid degradation of endogenous proteins by Trim-Away

et al. 2018

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This Protocol describes 'Trim-away', an approach for rapid protein depletion in different cell-types. TRIM21-mediated proteasomal degradation is induced by microinjection or electroporation of an antibody against the protein of interest. TWEET A new Protocol describing the detailed procedures for 'Trim-away', an approach for rapid protein depletion in oocytes, primary cells, and cultured cells @SchuhLab @mpi_bpc @MRC_LMB @CellBiol_MRCLMB COVER TEASER Rapid protein depletion in cells using 'Trim-away' Please indicate up to four primary research articles where the protocol has been used and/or developed. 1. Clift, D. et al. A method for the acute and rapid degradation of endogenous proteins. Cell. 171, 1692-1706 (2017).

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Section: Experimental Designmentioning

confidence: 99%

Acute and rapid degradation of endogenous proteins by Trim-Away

et al. 2018

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“…Extrapolating from this notion, an interesting hypothesis is that the nuclear deformation under mechanical stress during confined migration might influence the genome architecture and gene expression, and thereby affect the final fate and destination of migrating neurons. Emerging techniques for visualization of genome architecture in live cells in combination with high throughput genome sequencing analyses will offer greater opportunities to answer these questions (Miyanari et al, 2013;Natale et al, 2017;Conic et al, 2018).…”

Section: Resultsmentioning

confidence: 99%

Mechanical Regulation of Nuclear Translocation in Migratory Neurons

Nakazawa

Kengaku

2020

Front. Cell Dev. Biol.

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Neuronal migration is a critical step during the formation of functional neural circuits in the brain. Newborn neurons need to move across long distances from the germinal zone to their individual sites of function; during their migration, they must often squeeze their large, stiff nuclei, against strong mechanical stresses, through narrow spaces in developing brain tissue. Recent studies have clarified how actomyosin and microtubule motors generate mechanical forces in specific subcellular compartments and synergistically drive nuclear translocation in neurons. On the other hand, the mechanical properties of the surrounding tissues also contribute to their function as an adhesive support for cytoskeletal force transmission, while they also serve as a physical barrier to nuclear translocation. In this review, we discuss recent studies on nuclear migration in developing neurons, from both cell and mechanobiological viewpoints.

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“…With a pulse time from microseconds to a second, the delivery window for proteins is very short as the membrane pores immediately shrink and reseal when the voltage is switched off [44]. Nonetheless, various proteins including antibodies, CRISPR/ Cas9 RNA-protein complexes and α-synuclein have been delivered into cells by bulk electroporation [45][46][47][48][49].…”

Section: Bulk and Localized Electroporationmentioning

confidence: 99%

Methods for protein delivery into cells: from current approaches to future perspectives

Chau

Actis

Hewitt

2020

Biochemical Society Transactions

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The manipulation of cultured mammalian cells by the delivery of exogenous macromolecules is one of the cornerstones of experimental cell biology. Although the transfection of cells with DNA expressions constructs that encode proteins is routine and simple to perform, the direct delivery of proteins into cells has many advantages. For example, proteins can be chemically modified, assembled into defined complexes and subject to biophysical analyses prior to their delivery into cells. Here, we review new approaches to the injection and electroporation of proteins into cultured cells. In particular, we focus on how recent developments in nanoscale injection probes and localized electroporation devices enable proteins to be delivered whilst minimizing cellular damage. Moreover, we discuss how nanopore sensing may ultimately enable the quantification of protein delivery at single-molecule resolution.

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Imaging of native transcription factors and histone phosphorylation at high resolution in live cells

Cited by 36 publications

References 69 publications

Acute and rapid degradation of endogenous proteins by Trim-Away

Acute and rapid degradation of endogenous proteins by Trim-Away

Mechanical Regulation of Nuclear Translocation in Migratory Neurons

Methods for protein delivery into cells: from current approaches to future perspectives

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