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Sigma-Aldrich® Advanced Genomics

Efficient Generation of Gene-edited Mouse Models and Cell Lines Using Synthetic sgRNA

The CRISPR/Cas genome editing system has revolutionized nearly every aspect of the life science industry. Until recently, the most used formats for this technology have been plasmids, mRNA, or lentivirus. Synthetic single modified guide RNA (sgRNA) delivered pre-complexed with Cas nucleases reduces the limiting factors of the above formats such as off-target effects, low efficiency,…

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How to Validate a CRISPR Experiment  

Discover how to validate the success of your CRISPR gene editing experiment from determining successful delivery of CRISPR reagents into your cells to the confirmation of desired genetic and phenotype changes.

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Engineering Vero Cell Lines Using CRISPR to Increase Production of Viral Vaccines

Engineering Vero Cell Lines Using CRISPR to Increase Production of Viral Vaccines On Demand Webinar Dr. Benjamin Borgo Dr. Benjamin Borgo currently leads a team of forward-thinking product and service managers within Merck KGaA’s Genome Engineering and Modulation franchise. His first exposure to CRISPR-based genome editing was as a graduate student where he attempted to…

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CRISPR Nucleases: The Ultimate Guide To Selection

CRISPR nucleases continue to dominate the gene-editing landscape because they are powerful and easy-to-use gene-editing tools. Their rapid discovery and development, however, make choosing the right nuclease for your experiment difficult. This guide highlights the broad range of available CRISPR nucleases and distils the major factors you should consider when choosing one for your next…

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CRISPR Multiplexing Strategies: What’s The Choice?

Although multiplex CRISPR gene editing can be accomplished by simply introducing more than one gRNA to your target cells, there are many alternative — and more efficient — ways of achieving this goal. This article discusses these alternative CRISPR multiplexing strategies and highlights their potential caveats. Not sure whether multiplex CRISPR gene editing is right…

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Multiplex CRISPR Gene Editing: What’s It For?

The RNA-guided nature of CRISPR nucleases (e.g., Cas9, Cpf1) makes them highly amenable to multiplex applications, including multi-gene knockout and large-scale genomic modifications. This article — part 1 of a two-part series on multiplex CRISPR gene editing — discusses these applications and outlines practical considerations for multiplex CRISPR gene editing experiments. New to CRISPR gene…

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Dead Useful: CRISPR-Cas9 Epigenome Editing

Given the rapid pace with which genome editing technologies have been developed and adopted, it comes as no surprise that the original CRISPR-Cas9 system has been successfully modified by some very clever scientists. No longer are we limited to the ‘simple’ case of editing the genetic sequence of your biological system of choice, but there…

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Get Prepped: Nanopore Library Preparation Optimization

Nanopore is a relatively new sequencing platform and researchers are still trying to optimize the protocol for their own specific applications. In our lab, we work primarily with metagenomic samples and use the 1D sequencing kits. Over the past year, we have optimized this technique. To check the quality of the Nanopore library preparation we…

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CRISPR-Based Activation (CRISPRa) of Genes: A How-To Guide

The purpose of this article is to walk you through the steps needed to overexpress genes using CRISPR/Cas9-based activation (CRISPRa). A broader overview of this topic (including CRISPR-based repression) can be found here. CRISPR/Cas9 is more than a programmable nuclease. When stripped of its nuclease activity, it can activate and repress transcription, alter chromatin structure,…

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CRISPR-Cas9 Genome Editing: Weighing the Pros and Cons

Genome editing is a hugely powerful tool which can help you to address a multitude of questions in your research. However, it is not necessarily the best tool for the job in every situation. Below is a discussion of the main advantages and disadvantages associated using CRISPR-Cas9 for genome editing. The Pros It’s Simple to…

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What’s that Organism? Using DNA Barcoding for Species Identification

In both the lab and field, it is important to know what species we are working with. While morphological data has always been a tried and true method of identifying species, DNA barcoding allows us to identify species when we don’t have that option (e.g. if we don’t have enough of a specimen to identify…

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How to Confirm Your CRISPR-cas9 Genome Editing Was Successful

You have sweated at the lab bench and in the cell culture suite for weeks (or more likely months) to plan and optimize for successful genome editing. Finally, you’ve got the guide RNAs and the vectors into the cells. Yes! But your work is not yet finished. You can’t take for granted the fact that…

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