CRISPR/Cas9 bioluminescence-based assay for monitoring CFTR trafficking to the plasma membrane
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15640%2F24%3A73621987" target="_blank" >RIV/61989592:15640/24:73621987 - isvavai.cz</a>
Alternative codes found
RIV/61989592:15110/24:73621987
Result on the web
<a href="https://www.life-science-alliance.org/content/7/1/e202302045" target="_blank" >https://www.life-science-alliance.org/content/7/1/e202302045</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.26508/lsa.202302045" target="_blank" >10.26508/lsa.202302045</a>
Alternative languages
Result language
angličtina
Original language name
CRISPR/Cas9 bioluminescence-based assay for monitoring CFTR trafficking to the plasma membrane
Original language description
CFTR is a membrane protein that functions as an ion channel. Mutations that disrupt its biosynthesis, trafficking or function cause cystic fibrosis (CF). Here, we present a novel in vitro model system prepared using CRISPR/Cas9 genome editing with en-dogenously expressed WT-CFTR tagged with a HiBiT peptide. To enable the detection of CFTR in the plasma membrane of live cells, we inserted the HiBiT tag in the fourth extracellular loop of WT-CFTR. The 11-amino acid HiBiT tag binds with high affinity to a large inactive subunit (LgBiT), generating a reporter luciferase with bright luminescence. Nine homozygous clones with the HiBiT knock-in were identified from the 182 screened clones; two were genetically and functionally validated. In summary, this work describes the preparation and validation of a novel reporter cell line with the potential to be used as an ultimate building block for developing unique cellular CF models by CRISPR-mediated insertion of CF-causing mutations.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10602 - Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Life Science Alliance
ISSN
2575-1077
e-ISSN
2575-1077
Volume of the periodical
7
Issue of the periodical within the volume
1
Country of publishing house
US - UNITED STATES
Number of pages
15
Pages from-to
"e202302045"
UT code for WoS article
001104243100001
EID of the result in the Scopus database
2-s2.0-85176200316