Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference

In recent years, CRISPR interference (CRISPRi) technology of gene silencing has emerged as a promising alternative to RNA interference (RNAi) surpassing the latter in terms of efficiency and accuracy. Here, we describe the construction of a set of transposon vectors suitable for constitutive or tetr...

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Autores principales: Czarnek, Maria, Kochan, Jakub, Wawro, Mateusz, Myrczek, Rafał, Bereta, Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2023
Materias:
Original Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10471651/
https://www.ncbi.nlm.nih.gov/pubmed/36707469
http://dx.doi.org/10.1007/s12033-023-00675-5
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author Czarnek, Maria
Kochan, Jakub
Wawro, Mateusz
Myrczek, Rafał
Bereta, Joanna
author_facet Czarnek, Maria
Kochan, Jakub
Wawro, Mateusz
Myrczek, Rafał
Bereta, Joanna
author_sort Czarnek, Maria
collection PubMed
description In recent years, CRISPR interference (CRISPRi) technology of gene silencing has emerged as a promising alternative to RNA interference (RNAi) surpassing the latter in terms of efficiency and accuracy. Here, we describe the construction of a set of transposon vectors suitable for constitutive or tetracycline (doxycycline)-inducible silencing of genes of interest via CRISPRi method and conferring three different antibiotic resistances, using vectors available via Addgene repository. We have analyzed the performance of the new vectors in the silencing of mouse Adam10 and human lncRNA, NORAD. The empty vector variants can be used to efficiently silence any genes of interest.
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spelling pubmed-104716512023-09-02 Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference Czarnek, Maria Kochan, Jakub Wawro, Mateusz Myrczek, Rafał Bereta, Joanna Mol Biotechnol Original Paper In recent years, CRISPR interference (CRISPRi) technology of gene silencing has emerged as a promising alternative to RNA interference (RNAi) surpassing the latter in terms of efficiency and accuracy. Here, we describe the construction of a set of transposon vectors suitable for constitutive or tetracycline (doxycycline)-inducible silencing of genes of interest via CRISPRi method and conferring three different antibiotic resistances, using vectors available via Addgene repository. We have analyzed the performance of the new vectors in the silencing of mouse Adam10 and human lncRNA, NORAD. The empty vector variants can be used to efficiently silence any genes of interest. Springer US 2023-01-28 2023 /pmc/articles/PMC10471651/ /pubmed/36707469 http://dx.doi.org/10.1007/s12033-023-00675-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Paper
Czarnek, Maria
Kochan, Jakub
Wawro, Mateusz
Myrczek, Rafał
Bereta, Joanna
Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title_full Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title_fullStr Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title_full_unstemmed Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title_short Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference
title_sort construction of a set of novel transposon vectors for efficient silencing of protein and lncrna genes via crispr interference
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10471651/
https://www.ncbi.nlm.nih.gov/pubmed/36707469
http://dx.doi.org/10.1007/s12033-023-00675-5
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