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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

Cpf1: A New Tool for CRISPR Genome Editing

Posted by Mary Gearing on Oct 14, 2015 10:30:00 AM

This post was updated on Dec 5, 2017.

In 2015, Zetsche et al. added to the CRISPR toolbox with their characterization of two Cpf1 orthologs that display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members contain a RuvC-like endonuclease domain, but they lack Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than the two (tracrRNA and crRNA) needed by Cas9 for cleavage. In certain cases, Cpf1 may be better suited for genome editing than Cas9 - read on to learn more about Cpf1 and check out our CRISPR guide for a refresher on CRISPR/Cas9. 

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Topics: Genome Engineering, CRISPR

pSiM24: Simplifying Plant Genetic Engineering

Posted by Mary Gearing on Sep 29, 2015 10:30:00 AM

As previous blogs have noted, plants are an important foundation for life on Earth. Selective breeding methods have shaped the plants that we grow and eat, and genetic engineering will continue to improve plant nutrition, yield, and pest resistance. Much of plant genetic engineering revolves around Agrobacterium tumifaciens. Agrobacterium carries a “tumor-inducing” or Ti plasmid, which allows it to transfer genetic material into the host plant genome. Scientists have worked to optimize this system for gene transfer, studying the stability of modified Ti plasmids during plant infection, as well as plasmid yield during preparation in E. coli. Addgene depositor Indu Maiti has created a new and versatile binary Ti vector for both transient and stable gene expression applications in plants. This smaller, easily customizable vector functions in multiple species, including tobacco and Arabidopsis.

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Topics: Plasmid Technology, Plant Biology

Synthetic Photobiology: Optogenetics for E. coli

Posted by Mary Gearing on Sep 8, 2015 10:30:00 AM

As optogenetics turns 10 years old, it’s easy to forget that this technique isn’t limited to neuroscience. In fact, precise light-based control of biological processes is highly useful in other fields, including synthetic biology. Addgene depositors Christopher Voigt and Jeffrey Tabor have been working on making E. coli light responsive since 2005, when Tabor was working in Voigt's lab. Years later, these classic systems continue to be optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust.

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Topics: Optogenetics, Synthetic Biology

Plasmids 101: Golden Gate Cloning

Posted by Mary Gearing on Aug 27, 2015 10:30:00 AM

Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method has its own pluses and minuses, but Golden Gate cloning has been especially useful within both the synthetic biology and genome engineering fields. We’ll walk you through how to apply this precise and easy-to-use system to your cloning efforts.

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Topics: Plasmid How To, Synthetic Biology, Plasmids 101, Protocols, Techniques, Plasmid Cloning

Mapping the 4D nucleome with CRISPR/Cas9

Posted by Mary Gearing on Aug 11, 2015 10:30:00 AM

It seems that there’s a new CRISPR advance or technique published every week! One of the newest applications is a colorful system that uses fluorescently labeled Cas9 to label multiple genomic loci in live cells. While other systems can be used to label loci, such as fluorescence in situ hybridization (FISH) or fluorescently labeled TALEs, CRISPR/Cas9’s ease of use and ability to label live cells make this system truly advantageous. This new technique, developed in Thoru Pederson’s lab, brings us one step closer to mapping the 4D nucleome, the organization of the nucleus in space and time, and to understanding how nuclear organization varies across the life of a cell, or how organization may be altered in disease states.

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Topics: CRISPR, Imaging, Fluorescent Proteins

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