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It’s time to choose your own protein purification adventure. You want to purify your favorite protein (YFP). You have two options: 

Option #1: Affinity tag purification

You tag YFP and use an affinity column for purification. After binding YFP to the column, you wash several times to remove non-specific proteins, and then elute YFP. 

Option #2: Opto-Nanobodies (OptoNBs) purification

You skip adding a tag to YFP and instead use OptoNBs. You fill a column with OptoNB coated beads and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific proteins flow through. To elute YFP, you turn on the blue lights.

Which option do you choose? 

CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability to infiltrate cells. Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction; but these methods aren’t stealthy enough to gain access to some cells and tissues, such as human induced pluripotent stem cells (hiPSCs). Nanoblades, a new CRISPR delivery method developed by the Ricci Lab and the T. Ohlmann Lab, adds a covert tool to the CRISPR tool box. 

One of the biggest barriers to gene therapy treatment of central nervous system (CNS) diseases is the blood brain barrier (BBB). It’s the BBB’s job to block the entry of pathogens to the CNS, but this also stops viral vectors, such as adeno-associated virus (AAV), from reaching their targets.

When I started writing for the Addgene blog, I was focused on writing about new scientific techniques and cool plasmids. Creating graphics were usually the last thing I thought about when writing posts. Since then I’ve realized my figures are just as important, if not more important, than my writing. Initially I didn’t have access to professional-grade design software, like Adobe Illustrator, and I didn’t want to pay for these programs either. But with a little Googling and some trial and error, I found some free design software that let me create graphics that better communicated the science in my blog posts. This post highlights several of these free tools which will hopefully also help you communicate your science, whether it’s in presentations, manuscripts, or social media.

Direct reprogramming describes the process where differentiated cells are turned into a cell type of choice while bypassing the intermediate pluripotent state. Though a valuable tool for regenerative medicine, direct reprogramming is an inefficient process, with the majority of cells failing to develop the desired identity.

The development of single cell technologies, such as single cell RNA-seq (scRNA-seq), now allows scientists to identify the gene expression patterns unique to the few cells that successfully reprogram. Unfortunately scRNA-seq only captures the gene expression of cells at a single time point, making it difficult to investigate how early expression patterns influence reprogramming outcomes.