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CRISPR Antimicrobials

Posted by Mary Gearing on May 3, 2016 10:30:00 AM

The crisis of antibiotic resistance is upon us, and the world is unprepared. Each year in the United States, two million people will be infected by antibiotic resistant bacteria. Even when researchers develop new antibiotics, the onset of resistance is swift, as few as five years after introduction. Current antibiotic strategies are nonspecific - they harm any bacterial cell without a resistance gene, allowing resistant bacteria to multiply, spreading their resistance genes throughout the bacterial population. But what if we could specifically target only virulent or antibiotic resistant bacteria with a weapon that they’ll have less potential to become resistant to? CRISPR may provide a method for doing just that. While challenges remain in the delivery of these agents, CRISPR antimicrobials could become our newest line of defense against bacteria.

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Topics: Genome Engineering, CRISPR

Casilio: An Adaptive, Multitasking “CRISPR-OS”

Posted by Guest Blogger on Apr 26, 2016 10:30:00 AM

This post was contributed by guest bloggers Albert Cheng and Mark Wanner.

CRISPR-Cas9 offers a leap forward for genome editing, providing researchers with greatly enhanced accuracy, efficiency, and versatility. It has led to a tremendous acceleration of biomedical research, allowing for the modeling of human disease mutations in experimental model systems with previously unthinkable speed and precision. Furthermore, the ability to excise detrimental mutations and introduce functional sequences—as is being investigated with dystrophin/Duchenne muscular dystrophy at this time—is potentially transformative for human clinical care for some Mendelian diseases.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

Genome engineering using Cas9/gRNA Ribonucleoproteins (RNPs)

Posted by Joel McDade on Apr 21, 2016 10:30:00 AM

CRISPR has quickly become the preferred system for genome engineering due to its simplicity, as it requires only Cas9 and a guide RNA (gRNA).  Choosing the correct method to deliver both Cas9 and gRNAs to your target cells is absolutely critical as failure to adequately express either component will result in a failed experiment.  In our previous blog post entitled “CRISPR 101 - Mammalian Expression Systems and Delivery Methods” we provided a general overview of the most common ways in which you can deliver Cas9 and gRNAs to your target cells and discussed a few key advantages and disadvantages of each method. In this blog post, we will go into greater detail about why and how Cas9/gRNA Ribonucleoprotein complexes (Cas9 RNPs) are being used for genome engineering experiments and provide a general framework for getting started with Cas9 RNPs in your research.

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Topics: Genome Engineering, CRISPR, Techniques

Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

Posted by Guest Blogger on Apr 12, 2016 10:30:00 AM

This post was contributed by guest blogger Alan Wong.

The complexity of biological systems can hinder our attempts to study and engineer them, but what if we had a simple tool that allowed us to rapidly decode the complexity? The CombiGEM-CRISPR technology was developed with the goal of providing an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your favorite biological phenotype in a scalable way. This blog post will introduce you to this new technology, and guide you through the basics of CombiGEM-CRISPR experiments.

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Topics: Plasmid Technology, Genome Engineering, CRISPR

Optimizing Donor DNA for Enhanced CRISPR Genome Editing

Posted by Guest Blogger on Mar 24, 2016 10:30:00 AM

This post was contributed by guest blogger Chris Richardson, a Postdoctoral Researcher in Jacob Corn’s lab.

CRISPR-Cas9 (Cas9) is an RNA-guided nuclease that targets and cuts genomic DNA. The interplay between Cas9 (which causes the breaks) and host cell DNA repair factors (which repair those breaks) makes Cas9 extremely effective as a genome editing reagent. This interplay falls into two broad categories and thus, causes two types of editing outcomes: Cas9 breaks repaired by the non-homologous end-joining (NHEJ) pathway disrupt target gene sequences (thus inactivating genes), while breaks repaired by homology directed repair (HDR) pathways can modify the sequence of a gene (thus altering its function). HDR is crucial for certain applications, for example, correcting the allele that causes sickle cell anemia. However, HDR occurs much less frequently than NHEJ and the efficiency of these editing reactions is low. Understanding the biological cause of this repair bias is a fascinating (and yet unanswered) question. Our recent paper (Richardson et al 2016) revealed some of the biophysical parameters that can influence the HDR/NHEJ decision.

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Topics: Genome Engineering, CRISPR

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