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R Bodies: Membrane-Rupturing Microscopic Tools

Posted by Guest Blogger on Apr 14, 2016 10:30:00 AM

This post was contributed by guest blogger Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver. 

Most types of biological motion (whether endocytosis, vesicle trafficking, or muscle contractions) are produced by orchestrated movements of networks of proteins consuming molecular fuel sources. While the importance of understanding these complex processes can’t be overstated, we can also learn a lot from Nature’s simpler solutions to transmitting forces over long distances. For instance, how much force can be generated by conformational changes in proteins? How can information propagate through a structured material over a long distance? And can we understand such a structure well enough to engineer it to suit our purposes?

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Topics: Plasmid Technology, Synthetic Biology

Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

Posted by Guest Blogger on Apr 12, 2016 10:30:00 AM

This post was contributed by guest blogger Alan Wong.

The complexity of biological systems can hinder our attempts to study and engineer them, but what if we had a simple tool that allowed us to rapidly decode the complexity? The CombiGEM-CRISPR technology was developed with the goal of providing an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your favorite biological phenotype in a scalable way. This blog post will introduce you to this new technology, and guide you through the basics of CombiGEM-CRISPR experiments.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

Modulate the Activity of 17 Signaling Pathways with One Kit!

Posted by Kendall Morgan on Mar 22, 2016 10:30:00 AM

When cancers are treated with drugs designed to hit them right where it hurts, the effects are often remarkable but fleeting.

“What’s been shown by others is that, in a relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said Kris Wood of Duke University Medical Center. “While we do not yet have a comprehensive understanding of how cancers become resistant, an emerging theme is that they do so by activating signaling pathways controlling properties like growth, survival, and differentiation.”

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Topics: Plasmid Technology, Cancer, Plasmid Kits

Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

Posted by Brook Pyhtila on Mar 1, 2016 10:30:00 AM

Over the past decade, scientists have developed and fine tuned many different ways to clone DNA fragments which have provided appealing alternatives to restriction enzyme cloning. These newer technologies have become more and more common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning we once relied exclusively on. In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using Gibson assembly to put together DNA fragments.

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Topics: Plasmid Technology, Plasmids 101, Protocols, Plasmid Cloning

PITChing MMEJ as an Alternative Route for Gene Editing

Posted by Mary Gearing on Feb 23, 2016 10:30:00 AM

If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions or homology-directed repair (HDR) to create precise genome edits. But have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated end-joining)? MMEJ, a form of alternative end-joining, requires only very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh (Precise Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for complicated cloning of homology arms.

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Topics: Plasmid Technology, Genome Engineering, CRISPR, Techniques

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