Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains

We have generated Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains where CRISPR/Cas9 was used to endogenously tag the protein at either the C- or N-terminus and validated the functionality of the resulting tagged TDPT-1 proteins. We have found that both the N-terminally tagged ( wrmScarle...

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Detalles Bibliográficos
Autores principales: Bhandari, Nirajan, Pfeiffer, Shannon C., Jaramillo-Lambert, Aimee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2022
Materias:
New Finding
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9010224/
https://www.ncbi.nlm.nih.gov/pubmed/35622466
http://dx.doi.org/10.17912/micropub.biology.000540
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author Bhandari, Nirajan
Pfeiffer, Shannon C.
Jaramillo-Lambert, Aimee
author_facet Bhandari, Nirajan
Pfeiffer, Shannon C.
Jaramillo-Lambert, Aimee
author_sort Bhandari, Nirajan
collection PubMed
description We have generated Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains where CRISPR/Cas9 was used to endogenously tag the protein at either the C- or N-terminus and validated the functionality of the resulting tagged TDPT-1 proteins. We have found that both the N-terminally tagged ( wrmScarlet::tdpt-1) and C-terminally tagged ( tdpt-1::3xflag ) worm TDPT-1 does not affect embryonic viability compared to wild type. Using the N-terminally tagged wrmScarlet::tdpt-1 strain we show, for the first time, that TDPT-1 is expressed in nuclei of the germ line and the soma. Moreover, we validate the expression of TDPT-1 at the protein level using the C-terminally tagged ( tdpt-1::3xflag ) strain.
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spelling pubmed-90102242022-04-15 Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains Bhandari, Nirajan Pfeiffer, Shannon C. Jaramillo-Lambert, Aimee MicroPubl Biol New Finding We have generated Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains where CRISPR/Cas9 was used to endogenously tag the protein at either the C- or N-terminus and validated the functionality of the resulting tagged TDPT-1 proteins. We have found that both the N-terminally tagged ( wrmScarlet::tdpt-1) and C-terminally tagged ( tdpt-1::3xflag ) worm TDPT-1 does not affect embryonic viability compared to wild type. Using the N-terminally tagged wrmScarlet::tdpt-1 strain we show, for the first time, that TDPT-1 is expressed in nuclei of the germ line and the soma. Moreover, we validate the expression of TDPT-1 at the protein level using the C-terminally tagged ( tdpt-1::3xflag ) strain. Caltech Library 2022-03-17 /pmc/articles/PMC9010224/ /pubmed/35622466 http://dx.doi.org/10.17912/micropub.biology.000540 Text en Copyright: © 2022 by the authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle New Finding
Bhandari, Nirajan
Pfeiffer, Shannon C.
Jaramillo-Lambert, Aimee
Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title_full Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title_fullStr Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title_full_unstemmed Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title_short Characterization of N- and C-terminal endogenously tagged Tyrosyl-DNA phosphodiesterase 2 (TDPT-1) C. elegans strains
title_sort characterization of n- and c-terminal endogenously tagged tyrosyl-dna phosphodiesterase 2 (tdpt-1) c. elegans strains
topic New Finding
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9010224/
https://www.ncbi.nlm.nih.gov/pubmed/35622466
http://dx.doi.org/10.17912/micropub.biology.000540
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