Getting the most out of publicly available T-DNA insertion lines

In the course of several different projects, we came to realize that there is a significant amount of untappedpotential in the publicly available T-DNA insertion lines. In addition to the GABI-Kat lines, which were designedspecifically for activation tagging, lines from the SAIL and FLAGdb collectio...

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Main Authors: Ulker, B., Peiter, E., Dixon, D., Moffat, Caroline, Capper, R., Bouche, N., Edwards, R., Sanders, D., Knight, H., Knight, M.
Format: Journal Article
Published: Wiley 2008
Subjects:
Online Access:http://www.ncbi.nlm.nih.gov/pubmed/18644000
http://hdl.handle.net/20.500.11937/45695
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author Ulker, B.
Peiter, E.
Dixon, D.
Moffat, Caroline
Capper, R.
Bouche, N.
Edwards, R.
Sanders, D.
Knight, H.
Knight, M.
author_facet Ulker, B.
Peiter, E.
Dixon, D.
Moffat, Caroline
Capper, R.
Bouche, N.
Edwards, R.
Sanders, D.
Knight, H.
Knight, M.
author_sort Ulker, B.
building Curtin Institutional Repository
collection Online Access
description In the course of several different projects, we came to realize that there is a significant amount of untappedpotential in the publicly available T-DNA insertion lines. In addition to the GABI-Kat lines, which were designedspecifically for activation tagging, lines from the SAIL and FLAGdb collections are also useful for this purpose.As well as the 35S promoter chosen for activation tagging in GABI-Kat lines, we found that the 1¢2¢ bidirectionalpromoter is capable of activating expression of flanking genomic sequences in both GABI-Kat and SAIL lines.Thus these lines have added potential for activation tagging. We also show that these lines are capable ofgenerating antisense transcripts and so have the potential to be used for suppression (loss/reduction offunction) studies. By virtue of weak terminator sequences in some T-DNA constructs, transcript read-throughfrom selectable markers is also possible, which again has the potential to be exploited in activation/suppression studies. Finally, we show that, by selecting and characterizing lines in which the T-DNA insertionsare present specifically within introns of a target gene, an allelic series of mutants with varying levels ofreduced expression can be generated, due to differences in efficiency of intron splicing. Taken together, ouranalyses demonstrate that there is a wealth of untapped potential within existing insertion lines for studies ongene function, and the effective exploitation of these resources is discussed.
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institution Curtin University Malaysia
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spelling curtin-20.500.11937-456952017-02-28T01:41:48Z Getting the most out of publicly available T-DNA insertion lines Ulker, B. Peiter, E. Dixon, D. Moffat, Caroline Capper, R. Bouche, N. Edwards, R. Sanders, D. Knight, H. Knight, M. read-through antisense intron splicing activation tagging T-DNA 1¢2¢ promoter In the course of several different projects, we came to realize that there is a significant amount of untappedpotential in the publicly available T-DNA insertion lines. In addition to the GABI-Kat lines, which were designedspecifically for activation tagging, lines from the SAIL and FLAGdb collections are also useful for this purpose.As well as the 35S promoter chosen for activation tagging in GABI-Kat lines, we found that the 1¢2¢ bidirectionalpromoter is capable of activating expression of flanking genomic sequences in both GABI-Kat and SAIL lines.Thus these lines have added potential for activation tagging. We also show that these lines are capable ofgenerating antisense transcripts and so have the potential to be used for suppression (loss/reduction offunction) studies. By virtue of weak terminator sequences in some T-DNA constructs, transcript read-throughfrom selectable markers is also possible, which again has the potential to be exploited in activation/suppression studies. Finally, we show that, by selecting and characterizing lines in which the T-DNA insertionsare present specifically within introns of a target gene, an allelic series of mutants with varying levels ofreduced expression can be generated, due to differences in efficiency of intron splicing. Taken together, ouranalyses demonstrate that there is a wealth of untapped potential within existing insertion lines for studies ongene function, and the effective exploitation of these resources is discussed. 2008 Journal Article http://hdl.handle.net/20.500.11937/45695 http://www.ncbi.nlm.nih.gov/pubmed/18644000 Wiley restricted
spellingShingle read-through
antisense
intron splicing
activation tagging
T-DNA
1¢2¢ promoter
Ulker, B.
Peiter, E.
Dixon, D.
Moffat, Caroline
Capper, R.
Bouche, N.
Edwards, R.
Sanders, D.
Knight, H.
Knight, M.
Getting the most out of publicly available T-DNA insertion lines
title Getting the most out of publicly available T-DNA insertion lines
title_full Getting the most out of publicly available T-DNA insertion lines
title_fullStr Getting the most out of publicly available T-DNA insertion lines
title_full_unstemmed Getting the most out of publicly available T-DNA insertion lines
title_short Getting the most out of publicly available T-DNA insertion lines
title_sort getting the most out of publicly available t-dna insertion lines
topic read-through
antisense
intron splicing
activation tagging
T-DNA
1¢2¢ promoter
url http://www.ncbi.nlm.nih.gov/pubmed/18644000
http://hdl.handle.net/20.500.11937/45695