Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag

Antony Adamson

doi:10.1101/331496

Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag

Antony Adamson

FBMH Platform Sciences, Enabling Technologies & Infrastructure

Research output: Contribution to journal › Article

Abstract

Electron microscopy has been the “gold standard” for studying collagen networks but dynamic information on how cells synthesise the networks has been lacking. Live imaging methods have been unable to distinguish newly-synthesised fibrils from pre-existing fibrils and intracellular collagen. Here, we tagged endogenous collagen-I using CRISPR-Cas9 with photoswitchable Dendra2 and demonstrate live cells synthesising, migrating on, and interacting with, collagen fibrils. This strategy is applicable for other long half-life proteins.

Original language	English
Journal	bioRxiv
DOIs	https://doi.org/10.1101/331496
Publication status	Published - 15 Jun 2018

Access to Document

10.1101/331496

Cite this

@article{2b1e30723e5441fe991f3186c29f1a42,

title = "Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag",

abstract = "Electron microscopy has been the “gold standard” for studying collagen networks but dynamic information on how cells synthesise the networks has been lacking. Live imaging methods have been unable to distinguish newly-synthesised fibrils from pre-existing fibrils and intracellular collagen. Here, we tagged endogenous collagen-I using CRISPR-Cas9 with photoswitchable Dendra2 and demonstrate live cells synthesising, migrating on, and interacting with, collagen fibrils. This strategy is applicable for other long half-life proteins.",

author = "Antony Adamson",

year = "2018",

month = jun,

day = "15",

doi = "10.1101/331496",

language = "English",

journal = "bioRxiv",

issn = "2692-8205",

publisher = "Cold Spring Harbor Laboratory Press",

}

TY - JOUR

T1 - Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag

AU - Adamson, Antony

PY - 2018/6/15

Y1 - 2018/6/15

N2 - Electron microscopy has been the “gold standard” for studying collagen networks but dynamic information on how cells synthesise the networks has been lacking. Live imaging methods have been unable to distinguish newly-synthesised fibrils from pre-existing fibrils and intracellular collagen. Here, we tagged endogenous collagen-I using CRISPR-Cas9 with photoswitchable Dendra2 and demonstrate live cells synthesising, migrating on, and interacting with, collagen fibrils. This strategy is applicable for other long half-life proteins.

AB - Electron microscopy has been the “gold standard” for studying collagen networks but dynamic information on how cells synthesise the networks has been lacking. Live imaging methods have been unable to distinguish newly-synthesised fibrils from pre-existing fibrils and intracellular collagen. Here, we tagged endogenous collagen-I using CRISPR-Cas9 with photoswitchable Dendra2 and demonstrate live cells synthesising, migrating on, and interacting with, collagen fibrils. This strategy is applicable for other long half-life proteins.

UR - https://www.biorxiv.org/content/early/2018/06/02/331496

U2 - 10.1101/331496

DO - 10.1101/331496

M3 - Article

JO - bioRxiv

JF - bioRxiv

SN - 2692-8205

ER -

Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag

Abstract

Access to Document

Other files and links

Fingerprint

Cite this