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Quick, Versatile Plant Transgenesis with GreenGate Plasmids

Posted by Kendall Morgan on Jan 23, 2014 10:23:00 AM

A few years ago, researchers introduced what’s been called a major breakthrough in cloning technology with the Golden Gate method. For the first time, it was possible to quickly and efficiently assemble a large number of building blocks with just two or three inexpensive enzymes. Now, a new toolkit - aptly named GreenGate – offers all of those same advantages to researchers working in plant model systems.

“The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann of Heidelberg University. “What is different is the plasmids you use to bring this into your target organism. We have designed a Golden Gate system based on the daily needs of an advanced molecular plant science lab.”

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Topics: Plasmid Technology, Genome Engineering, Synthetic Biology, Plasmid Kits

Using CRISPR/Cas9 to Edit Disease Out of the Genome

Posted by Kendall Morgan on Jan 7, 2014 10:45:00 AM

There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. Now two studies reported in Cell Stem Cell last month show that this tool - already so useful in the laboratory - might also find its way to the clinic.

A team led by Jinsong Li from the Chinese Academy of Sciences found that mice with a dominant mutation in a gene that causes cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA targeting the mutant allele. An independent team led by Hans Clevers at Hubrecht Institute in The Netherlands used the CRISPR/Cas9 genome editing system to correct the cystic fibrosis transmembrane conductor receptor (CFTR) by homologous recombination in cultured intestinal stem cells of patients with cystic fibrosis.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

Kiran Musunuru on the Newest TALEN Genome-Editing System

Posted by Kendall Morgan on Dec 10, 2013 10:13:00 AM

The goal of Kiran Musunuru's lab in Harvard's Department of Stem Cell and Regenerative Biology is to understand the basis for cardiovascular and metabolic human diseases. They do that by studying patients to uncover new gene variants associated with conditions of interest, then studying those variants in model systems: either human cells or mice.

In a recent issue of Cell Stem Cell, Musunuru, Chad Cowan and their colleagues describe a much more efficient tool for doing that disease modeling work in human pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit consisting of 834 plasmids. The researchers showed they could use their TALEN kit to quickly and efficiently generate human stem cells edited to carry mutant versions of 15 different disease-associated genes.

Addgene spoke to Musunuru about how the new kit works, the research the kit now makes possible, and how it compares to his CRISPR/Cas9 system.

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Topics: Plasmid Technology, Genome Engineering, Interview, Investigator Feature, Plasmid Kits

History of CRISPR Cas - A tale of survival and evolution

Posted by Margo R. Monroe on Nov 25, 2013 5:12:00 PM

All organisms share an innate goal to survive. This past year, scientists hijacked survival tactics of prokaryotes to deliver the technological biological blockbuster known as the CRISPR (clustered regularly interspaced palindromic repeats) Cas (CRISPR associated genes) system. This popular genome engineering tool offers flexibility, multiplexibility, and ease of use. In order for this technology to survive the diverse demands of the biotech field, let's look at how prokaryotes originally utilized CRISPR/Cas as a powerful and adaptive defense strategy against life-threatening viruses.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

Let There Be LITE Plasmids

Posted by Kendall Morgan on Nov 21, 2013 10:56:00 AM

When neuroscience graduate student Silvana Konermann first entered Feng Zhang’s lab at MIT, the use of customizable DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering applications was still very new. Now, Konermann and her colleagues including Zhang and Mark Brigham have taken the technology to another level with the addition of two light-sensitive ingredients - CRY2 and CIB1 – which they borrowed from Arabidopsis thaliana. The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off, literally at the flip of a (blue) light switch.

Zhang’s team devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living brain tissue. “The brain is such an amazing and dynamic organ,” Konermann explained. “It helps us adapt to influences in our environments. We are able to react and learn. All of this requires genes to be regulated dynamically.”

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Topics: Plasmid Technology, Genome Engineering, Optogenetics

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