Gene Synthesis
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Rescooped by Elizabeth Jones from Plant Biology Teaching Resources (Higher Education)
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Nature Biotech: Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease (2013)

Nature Biotech: Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease (2013) | Gene Synthesis | Scoop.it

http://www.nature.com/nbt/journal/v31/n8/full/nbt.2655.html

 

Sustainable intensification of crop production is essential to ensure food demand is matched by supply as the human population continues to increase. This will require high-yielding crop varieties that can be grown sustainably with fewer inputs on less land. Both plant breeding and genetic modification (GM) methods make valuable contributions to varietal improvement, but targeted genome engineering promises to be critical to elevating future yields. Most such methods require targeting DNA breaks to defined locations followed by either nonhomologous end joining (NHEJ) or homologous recombination. Zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be engineered to create such breaks, but these systems require two different DNA binding proteins flanking a sequence of interest, each with a C-terminal FokI nuclease module. We report here that the bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR) system, comprising a CRISPR-associated (Cas)9 protein and an engineered single guide RNA (sgRNA) that specifies a targeted nucleic acid sequence, is applicable to plants to induce mutations at defined loci.


Via Kamoun Lab @ TSL, Mary Williams
Elizabeth Jones's insight:

CRISPR-Cas9 is hot right now, and it's being used in several different ways: for genome-editing in cells of various species (here they're showing it works in plants),  or to create heritable changes (i.e. new transgenic animal lines), and also as a transcription factor to regulate gene expression without altering the nuclear DNA. 

 

Constructs that use Cas9 and synthetic guide RNA (sgRNA) can be customized to target any gene of interest, and can produce insertions, deletions, or incorportation of variant sequences into genomic DNA. Customized constructs can be ordered from gene synthesis suppliers such as GenScript, and delivered much more quickly and cheaply than they can be produced in a typical lab using traditional molecular cloning techniques. 

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Scooped by Elizabeth Jones
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Efficient genome editing in Caenorhabditis elegans by CRISPR ...

Efficient genome editing in Caenorhabditis elegans by CRISPR ... | Gene Synthesis | Scoop.it
Abstract. Cas9 is an RNA-guided double-stranded DNA nuclease that participates in clustered regularly interspaced short palindromic repeats (CRISPR)-mediated adaptive immunity in prokaryotes. CRISPR–Cas9 has ...
Elizabeth Jones's insight:

CRISPR-Cas9 is hot right now, and it's being used in several different ways: for genome-editing in cells of various species or to create heritable changes (i.e. new transgenic lines), and also as a transcription factor to regulate gene expression without altering the nuclear DNA. 

 

Constructs that use Cas9 and synthetic guide RNA (sgRNA) can be customized to target any gene of interest, and can produce insertions, deletions, or incorportation of variant sequences into genomic DNA. Customized constructs can be ordered from gene synthesis suppliers such as GenScript, and delivered much more quickly and cheaply than they can be produced in a typical lab using traditional molecular cloning techniques. 

more...
No comment yet.