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Fluorescent proteins have enabled scientists to pursue creative research avenues previously unavailable to them. With these tools it’s now easy to monitor protein expression, localization, and protein-protein interactions. Beyond these common applications, researchers are finding new ways to apply fluorescent proteins everyday. 

While much of CRISPR research has focused on genome editing, numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific DNA sequence via a targeting gRNA. But dCas9 does not cleave the DNA. Much of the research using dCas9 has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64 (Gilbert et al., 2013), or repression of transcription through binding a promoter region to inhibit association of transcriptional activators (Qi et al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation of a genomic region of interest targeted by a gRNA.

In this quarterly blog series, we’ll highlight a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more in-depth, but we hope that this blog series will help you find new CRISPR tools for your research!

This time:

• E. coli genome-wide CRISPR inhibition
• Covalent tethering of DNA template to Cas9
• SECURE base editors
• Nme2Cas9
• CRISPR interference in Candida albicans

So you’ve done the research, gathered up your data into an exciting story, and are ready to present your findings at a conference. But what you get out of a conference depends on what you put into it before, during, and after the meeting. Let’s break it down into the following: the elevator pitch, talks, the poster session, networking, and social media.

Apoptosis or “programmed cell death" plays a pivotal role in an array of biological processes including development, the immune system, and cell turnover. Apoptosis is a highly controlled process that is triggered by internal and external signals such as developmental cues and DNA damage. These signals activate a cascade of caspases, protease enzymes that cleave proteins. Executioner caspases are activated last in the cascade and are responsible for the degradation of over 600 cellular components ultimately leading to cell fragmentation and death (Elmore, 2007).