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Mary Gearing

Mary Gearing is a Scientist at Addgene. She got her start as a Science Communications Intern writing for the Addgene blog and website. As a full-time Addgenie, she still enjoys blogging about CRISPR and other cool plasmids!

Recent Posts

CRISPR 101: Multiplex expression of gRNAs

Posted by Mary Gearing on Jan 28, 2016 10:50:00 AM

This post was updated on Dec 5, 2017.

CRISPR makes it easy to target multiple loci - a concept called multiplexing. Since CRISPR is such a robust system, editing or labeling efficiency doesn’t usually change when you add multiple gRNAs. Sound good? Addgene has many tools to help you multiplex - we’ll use mammalian plasmids to introduce you to some of your potential options and cloning methods, but please scroll down for plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!

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Topics: Genome Engineering, CRISPR, CRISPR 101, Plasmid Kits

Treating Muscular Dystrophy with CRISPR Gene Editing

Posted by Mary Gearing on Jan 26, 2016 10:30:00 AM

Having seen CRISPR’s success in basic research, researchers are eager to apply it in a clinical setting. CRISPR is often used for animal germline modification, to repair or add in disease-causing mutations, but, until recently it hadn’t been used to treat disease postnatally. Now, three papers published concurrently in Science have shown CRISPR can treat a genetic disease in a postnatal mouse model, an important proof of concept for future preclinical and clinical work.

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Topics: CRISPR

Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

Posted by Mary Gearing on Dec 17, 2015 10:30:00 AM

If cloning methods had personalities, SLIC (sequence- and ligation-independent cloning) would be a true rebel. Not only does this system not use site-specific recombination, it also doesn’t require a ligation step! Based on the robust system of homologous recombination found in E. coli, SLIC is a cheap, standardized, and rapid multi-part DNA assembly method - read on to learn how to use it in your research.

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Topics: Plasmid How To, Plasmids 101, Protocols, Plasmid Cloning

Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

Posted by Mary Gearing on Nov 24, 2015 10:30:00 AM

At Addgene, we love GFP, and we’re always excited when depositors find new ways to make this workhorse protein even more useful! From FPs optimized for oxidizing environments to photoconvertible variants, it seems like GFP is always learning new things. Now, work from Connie Cepko’s lab allow researchers to activate transcription or Cre recombinase activity only in the presence of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time and money needed to develop new lines.

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Topics: Plasmid Technology, Synthetic Biology, Fluorescent Proteins, Cre-lox

Inntags: Innovative Protein Epitope Tagging

Posted by Mary Gearing on Nov 10, 2015 10:30:00 AM

First described in the 1980s, protein tags are now one of the most useful items in a scientist’s toolbox. As we’ve covered in Plasmids 101, tags can help you determine localization of a protein of interest, purify it, or determine its expression level without the need for a custom antibody. There is one major caveat - a tag may interfere with protein localization and/or function, so each tagged protein must be tested carefully to ensure it retains the attributes of the native protein. Since this process takes a lot of time and energy, Martí Aldea and collaborators have created a set of “innocuous tags” (inntags) less likely to alter a protein’s properties.

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Topics: Hot Plasmids, Plasmid Elements

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