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Rescooped by Lonicera.Gu from Plant Biology Teaching Resources (Higher Education)
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Three in Development on pattern formation and polarity

Three in Development on pattern formation and polarity | New collect | Scoop.it

If you love patterning like I love patterning, you might be interested in these three recent articles in Development.

 

Here's a Hypothesis:

 

"An intracellular partitioning-based framework for tissue cell polarity in plants and animals" http://dev.biologists.org/content/140/10/2061.abstract

 

"We propose that a fundamental building block for tissue cell polarity is the process of intracellular partitioning,which can establish individual cell polarity in the absence of asymmetric cues."

 

 

And another Hypothesis:

 

"Polar auxin transport: models and mechanisms"

http://dev.biologists.org/content/140/11/2253.abstract

 

"Here we propose a new mathematical framework for the analysis of polar auxin transport and present a detailed mathematical analysis of published models."

 

And a Open Access research article on leaf polarity, running title

"ARF3 is a direct target of AS1"

http://dev.biologists.org/content/140/9/1958.short

(Currently one of the most-read articles in Development)


Via Mary Williams
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Rescooped by Lonicera.Gu from TAL effector science
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A rapid assay to quantify the cleavage efficiency of custom-designed nucleases in planta - Plant Mol. Biol.

A rapid assay to quantify the cleavage efficiency of custom-designed nucleases in planta - Plant Mol. Biol. | New collect | Scoop.it

(via T. Lahaye, thanks...cant beat his speed :))

Johnson et al, 2013

Custom-designed nucleases are a promising technology for genome editing through the catalysis of double-strand DNA breaks within target loci and subsequent repair by the host cell, which can result in targeted mutagenesis or gene replacement. Implementing this new technology requires a rapid means to determine the cleavage efficiency of these custom-designed proteins in planta. Here we present such an assay that is based on cleavage-dependent luciferase gene correction as part of a transient dual-luciferase® reporter (Promega) expression system. This assay consists of co-infiltrating Nicotiana benthamiana leaves with two Agrobacterium tumefaciens strains: one contains the target sequence embedded within a luciferase reporter gene and the second strain contains the custom-designed nuclease gene(s). We compared repair following site-specific nuclease digestion through non-homologous DNA end-joining, as opposed to single strand DNA annealing, as a means to restore an out-of-frame luciferase gene cleavage-reporter construct. We show, using luminometer measurements and bioluminescence imaging, that the assay for non-homologous end-joining is sensitive, quantitative, reproducible and rapid in estimating custom-designed nucleases’ cleavage efficiency. We detected cleavage by two out of three transcription activator-like effector nucleases that we custom-designed for targets in the Arabidopsis CRUCIFERIN3 gene, and we compared with the well-established ‘QQR’ zinc-finger nuclease. The assay we report requires only standard equipment and basic plant molecular biology techniques, and it can be carried out within a few days. Different types of custom-designed nucleases can be preliminarily tested in our assay system before their downstream application in plant genome editing.


Via dromius
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