CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics
Single-molecule localization microscopy (SMLM) reports on protein organization in cells with near-molecular resolution and in combination with stoichiometric labeling enables protein counting. Fluorescent proteins allow stoichiometric labeling of cellular proteins; however, most methods either lead...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
2021
|
| In: |
iScience
Year: 2020, Volume: 24, Issue: 1 |
| ISSN: | 2589-0042 |
| DOI: | 10.1016/j.isci.2020.101895 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.isci.2020.101895 Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S2589004220310920 
|
| Author Notes: | Tim N. Baldering, Christos Karathanasis, Marie-Lena I.E. Harwardt, Petra Freund, Matthias Meurer, Johanna V. Rahm, Michael Knop, Marina S. Dietz, and Mike Heilemann |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1753176697 | ||
| 003 | DE-627 | ||
| 005 | 20241230010204.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 210406r20212020xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.isci.2020.101895 |2 doi | |
| 035 | |a (DE-627)1753176697 | ||
| 035 | |a (DE-599)KXP1753176697 | ||
| 035 | |a (OCoLC)1341403453 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 32 |2 sdnb | ||
| 100 | 1 | |a Baldering, Tim N. |d 1992- |e VerfasserIn |0 (DE-588)1230905413 |0 (DE-627)1753273226 |4 aut | |
| 245 | 1 | 0 | |a CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics |c Tim N. Baldering, Christos Karathanasis, Marie-Lena I.E. Harwardt, Petra Freund, Matthias Meurer, Johanna V. Rahm, Michael Knop, Marina S. Dietz, and Mike Heilemann |
| 246 | 3 | 0 | |a twelve |
| 264 | 1 | |c 2021 | |
| 300 | |a 20 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Available online 7 December 2020 | ||
| 500 | |a Supplemental information Seite 11-20 | ||
| 500 | |a Gesehen am 07.04.2021 | ||
| 520 | |a Single-molecule localization microscopy (SMLM) reports on protein organization in cells with near-molecular resolution and in combination with stoichiometric labeling enables protein counting. Fluorescent proteins allow stoichiometric labeling of cellular proteins; however, most methods either lead to overexpression or are complex and time demanding. We introduce CRISPR/Cas12a for simple and efficient tagging of endogenous proteins with a photoactivatable protein for quantitative SMLM and single-particle tracking. We constructed a HEK293T cell line with the receptor tyrosine kinase MET tagged with mEos4b and demonstrate full functionality. We determine the oligomeric state of MET with quantitative SMLM and find a reorganization from monomeric to dimeric MET upon ligand stimulation. In addition, we measured the mobility of single MET receptors in vivo in resting and ligand-treated cells. The combination of CRISPR/Cas12a-assisted endogenous protein labeling and super-resolution microscopy represents a powerful tool for cell biological research with molecular resolution. | ||
| 534 | |c 2020 | ||
| 650 | 4 | |a Biochemistry | |
| 650 | 4 | |a Biological Sciences Research Methodologies | |
| 650 | 4 | |a Biophysics | |
| 650 | 4 | |a Molecular Biology | |
| 700 | 1 | |8 1\p |a Karathanasis, Christos |d 1989- |e VerfasserIn |0 (DE-588)116915946X |0 (DE-627)1032791977 |0 (DE-576)511946635 |4 aut | |
| 700 | 1 | |a Harwardt, Marie-Lena I. E. |e VerfasserIn |4 aut | |
| 700 | 1 | |a Freund, Petra |e VerfasserIn |4 aut | |
| 700 | 1 | |a Meurer, Matthias |e VerfasserIn |0 (DE-588)1064186963 |0 (DE-627)813075610 |0 (DE-576)423831542 |4 aut | |
| 700 | 1 | |a Rahm, Johanna V. |e VerfasserIn |4 aut | |
| 700 | 1 | |a Knop, Michael |e VerfasserIn |0 (DE-588)1064187552 |0 (DE-627)813076749 |0 (DE-576)423832107 |4 aut | |
| 700 | 1 | |a Dietz, Marina S. |e VerfasserIn |4 aut | |
| 700 | 1 | |8 2\p |a Heilemann, Mike |e VerfasserIn |0 (DE-588)1096549891 |0 (DE-627)856929999 |0 (DE-576)33572535X |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t iScience |d Amsterdam : Elsevier, 2018 |g 24(2021), 1, Artikel-ID 101895 |h Online-Ressource |w (DE-627)1019532106 |w (DE-600)2927064-9 |w (DE-576)502115858 |x 2589-0042 |7 nnas |a CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics |
| 773 | 1 | 8 | |g volume:24 |g year:2021 |g number:1 |g elocationid:101895 |g extent:20 |a CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics |
| 856 | 4 | 0 | |u https://doi.org/10.1016/j.isci.2020.101895 |x Verlag |x Resolving-System |z lizenzpflichtig |3 Volltext |
| 856 | 4 | 0 | |u https://www.sciencedirect.com/science/article/pii/S2589004220310920 |x Verlag |z lizenzpflichtig |3 Volltext |
| 883 | |8 1\p |a cgwrk |d 20241001 |q DE-101 |u https://d-nb.info/provenance/plan#cgwrk | ||
| 883 | |8 2\p |a cgwrk |d 20241001 |q DE-101 |u https://d-nb.info/provenance/plan#cgwrk | ||
| 951 | |a AR | ||
| 992 | |a 20210406 | ||
| 993 | |a Article | ||
| 994 | |a 2021 | ||
| 998 | |g 1064187552 |a Knop, Michael |m 1064187552:Knop, Michael |d 700000 |d 706000 |e 700000PK1064187552 |e 706000PK1064187552 |k 0/700000/ |k 1/700000/706000/ |p 7 | ||
| 998 | |g 1064186963 |a Meurer, Matthias |m 1064186963:Meurer, Matthias |d 700000 |d 706000 |e 700000PM1064186963 |e 706000PM1064186963 |k 0/700000/ |k 1/700000/706000/ |p 5 | ||
| 999 | |a KXP-PPN1753176697 |e 3902161736 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"id":{"doi":["10.1016/j.isci.2020.101895"],"eki":["1753176697"]},"language":["eng"],"type":{"bibl":"article-journal","media":"Online-Ressource"},"origin":[{"dateIssuedKey":"2021","dateIssuedDisp":"2021"}],"title":[{"title":"CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics","title_sort":"CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamics"}],"person":[{"given":"Tim N.","display":"Baldering, Tim N.","family":"Baldering","role":"aut"},{"role":"aut","display":"Karathanasis, Christos","family":"Karathanasis","given":"Christos"},{"role":"aut","given":"Marie-Lena I. E.","family":"Harwardt","display":"Harwardt, Marie-Lena I. E."},{"given":"Petra","family":"Freund","display":"Freund, Petra","role":"aut"},{"given":"Matthias","display":"Meurer, Matthias","family":"Meurer","role":"aut"},{"given":"Johanna V.","display":"Rahm, Johanna V.","family":"Rahm","role":"aut"},{"display":"Knop, Michael","family":"Knop","given":"Michael","role":"aut"},{"role":"aut","family":"Dietz","display":"Dietz, Marina S.","given":"Marina S."},{"role":"aut","given":"Mike","family":"Heilemann","display":"Heilemann, Mike"}],"physDesc":[{"extent":"20 S."}],"relHost":[{"disp":"CRISPR/Cas12a-mediated labeling of MET receptor enables quantitative single-molecule imaging of endogenous protein organization and dynamicsiScience","pubHistory":["Volume 1 (March 23, 2018)-"],"note":["Gesehen am 11.09.2018"],"recId":"1019532106","id":{"issn":["2589-0042"],"zdb":["2927064-9"],"eki":["1019532106"]},"type":{"bibl":"periodical","media":"Online-Ressource"},"language":["eng"],"origin":[{"publisher":"Elsevier","publisherPlace":"Amsterdam ; Boston ; London ; New York ; Oxford ; Paris ; Philadelphia ; San Diego ; St. Louis","dateIssuedDisp":"[2018]-"}],"part":{"volume":"24","year":"2021","extent":"20","text":"24(2021), 1, Artikel-ID 101895","issue":"1"},"physDesc":[{"extent":"Online-Ressource"}],"title":[{"title":"iScience","title_sort":"iScience"}]}],"name":{"displayForm":["Tim N. Baldering, Christos Karathanasis, Marie-Lena I.E. Harwardt, Petra Freund, Matthias Meurer, Johanna V. Rahm, Michael Knop, Marina S. Dietz, and Mike Heilemann"]},"recId":"1753176697","note":["Available online 7 December 2020","Supplemental information Seite 11-20","Gesehen am 07.04.2021"]} | ||
| SRT | |a BALDERINGTCRISPRCAS12021 | ||