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Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

Posted by Guest Blogger on May 11, 2017 10:30:00 AM

This post was contributed  by Jesse S. Boehm, the Associate Director of the Cancer Program at the Broad Institute of Harvard and MIT.

The notion of cancer precision medicine seems so simple! Take a patient’s tumor sample, use cutting edge genomic technologies to map the mutations that are present, and use prior knowledge (data connecting each genotype with vulnerabilities) to design a therapeutic strategy that works.

But, those darn cancers have revealed many tricks up their sleeves and most patients still don’t benefit from this approach. One central bottleneck is that most recurrently mutated cancer genes are rare and most of the individual variants found in tumors are exceedingly rare. As a result, how most of these “variants of unknown significance” (sometimes called “VUS”) function is unknown. How can we make a decision for each patient if the majority of information on each cancer clinical sequencing report includes rare variants that haven’t been characterized?

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Topics: Cancer

Multiplex Genome Editing with CRISPR-Cpf1

Posted by Beth Kenkel on May 9, 2017 10:12:15 AM

There’s a new development for CRISPR-Cpf1 genome editing!  A recent paper from Feng Zhang's lab describes how to use Cpf1 for multiplex genome editing.  For a few reasons, Cpf1 is a simplified system for editing multiple targets compared to Cas9.  Read on to learn more about Cpf1 multiplexing.  For an in-depth review of Cpf1, check out this blog post or see Addgene's CRISPR guide page for a review of Cas9.  For a brief comparison of Cpf1 vs. Cas9, see the table below.

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Topics: CRISPR

Plasmids 101: Fluorescent Protein Timers

Posted by Tyler Ford on May 4, 2017 10:30:00 AM

Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at them under a microscope, fractionate samples to isolate particular organelles and their contents, or perform in situ hybridization experiments. In many cases fluorescent proteins have usurped old methods or complemented them in ways that make them much easier. A special class of FPs, the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers can look at a single image of a cell and understand how protein localization changes over time.
Download the Fluorescent Proteins 101 eBook
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Topics: Plasmids 101, Fluorescent Proteins

How to Design Your gRNA for CRISPR Genome Editing

Posted by Guest Blogger on May 3, 2017 11:00:00 AM

This Post was updated on May 3, 2017 with additional information and resources. 

This post was contributed by guest blogger, Addgene Advisory Board member, and Associate Director of the Genetic Perturbation Platform at the Broad Institute, John Doench.

CRISPR technology has made it easier than ever both to engineer specific DNA edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post will discuss differences between these approaches, as well as provide updates on how best to design gRNAs. You can also find validated gRNAs for your next experiment in Addgene's Validated gRNA Sequence Datatable.

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Topics: Plasmid How To, Genome Engineering, Lab Tips, CRISPR

Tips for Getting a Faculty Position

Posted by Guest Blogger on May 2, 2017 10:30:00 AM

This post was contributed by guest blogger Erik Snapp, Director of Graduate and Postdoctoral Programs at Janelia Research Campus.

Eight years ago, I decided to write a "how to" manual on applying for faculty positions in biomedical science. My motivation was to share my experiences from my own job search and my time on faculty search committees. Having successfully navigated the trials and tribulations of the process, I’ve provided guidance and mentoring to several people that found my insights helpful. All went on to get faculty positions at top state colleges, private universities, and medical schools.

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Topics: Career

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