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A Rice Stowaway MITE for Gene Transfer in Yeast

Miniature inverted repeat transposable elements (MITEs) lack protein coding capacity and often share very limited sequence similarity with potential autonomous elements. Their capability of efficient transposition and dramatic amplification led to the proposition that MITEs are an untapped rich sour...

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Autores principales: Fattash, Isam, Bhardwaj, Priyanka, Hui, Caleb, Yang, Guojun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3660474/
https://www.ncbi.nlm.nih.gov/pubmed/23704977
http://dx.doi.org/10.1371/journal.pone.0064135
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author Fattash, Isam
Bhardwaj, Priyanka
Hui, Caleb
Yang, Guojun
author_facet Fattash, Isam
Bhardwaj, Priyanka
Hui, Caleb
Yang, Guojun
author_sort Fattash, Isam
collection PubMed
description Miniature inverted repeat transposable elements (MITEs) lack protein coding capacity and often share very limited sequence similarity with potential autonomous elements. Their capability of efficient transposition and dramatic amplification led to the proposition that MITEs are an untapped rich source of materials for transposable element (TE) based genetic tools. To test the concept of using MITE sequence in gene transfer, a rice Stowaway MITE previously shown to excise efficiently in yeast was engineered to carry cargo genes (neo and gfp) for delivery into the budding yeast genome. Efficient excision of the cargo gene cassettes was observed even though the excision frequency generally decreases with the increase of the cargo sizes. Excised elements insert into new genomic loci efficiently, with about 65% of the obtained insertion sites located in genes. Elements at the primary insertion sites can be remobilized, frequently resulting in copy number increase of the element. Surprisingly, the orientation of a cargo gene (neo) on a construct bearing dual reporter genes (gfp and neo) was found to have a dramatic effect on transposition frequency. These results demonstrated the concept that MITE sequences can be useful in engineering genetic tools to deliver cargo genes into eukaryotic genomes.
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spelling pubmed-36604742013-05-23 A Rice Stowaway MITE for Gene Transfer in Yeast Fattash, Isam Bhardwaj, Priyanka Hui, Caleb Yang, Guojun PLoS One Research Article Miniature inverted repeat transposable elements (MITEs) lack protein coding capacity and often share very limited sequence similarity with potential autonomous elements. Their capability of efficient transposition and dramatic amplification led to the proposition that MITEs are an untapped rich source of materials for transposable element (TE) based genetic tools. To test the concept of using MITE sequence in gene transfer, a rice Stowaway MITE previously shown to excise efficiently in yeast was engineered to carry cargo genes (neo and gfp) for delivery into the budding yeast genome. Efficient excision of the cargo gene cassettes was observed even though the excision frequency generally decreases with the increase of the cargo sizes. Excised elements insert into new genomic loci efficiently, with about 65% of the obtained insertion sites located in genes. Elements at the primary insertion sites can be remobilized, frequently resulting in copy number increase of the element. Surprisingly, the orientation of a cargo gene (neo) on a construct bearing dual reporter genes (gfp and neo) was found to have a dramatic effect on transposition frequency. These results demonstrated the concept that MITE sequences can be useful in engineering genetic tools to deliver cargo genes into eukaryotic genomes. Public Library of Science 2013-05-21 /pmc/articles/PMC3660474/ /pubmed/23704977 http://dx.doi.org/10.1371/journal.pone.0064135 Text en © 2013 Fattash et al http://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 properly credited.
spellingShingle Research Article
Fattash, Isam
Bhardwaj, Priyanka
Hui, Caleb
Yang, Guojun
A Rice Stowaway MITE for Gene Transfer in Yeast
title A Rice Stowaway MITE for Gene Transfer in Yeast
title_full A Rice Stowaway MITE for Gene Transfer in Yeast
title_fullStr A Rice Stowaway MITE for Gene Transfer in Yeast
title_full_unstemmed A Rice Stowaway MITE for Gene Transfer in Yeast
title_short A Rice Stowaway MITE for Gene Transfer in Yeast
title_sort rice stowaway mite for gene transfer in yeast
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3660474/
https://www.ncbi.nlm.nih.gov/pubmed/23704977
http://dx.doi.org/10.1371/journal.pone.0064135
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