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Harnessing Bacterial Toxins for Allelic Exchange

Posted by Guest Blogger on Aug 15, 2019 8:30:02 AM

This post was contributed by Jacob Lazarus, a postdoctoral researcher at Harvard.

There’s an astounding number of ways to create chromosomal mutations in bacteria, so many that it may be difficult to decide which path to take. A quick and easy way to introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes. However, there are ways to create scarless mutations, ones that don’t leave any undesired scars in the chromosome.

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Topics: Genome Editing, Plasmids

Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

Posted by Guest Blogger on Aug 13, 2019 8:58:54 AM

This post was contributed by Katherine Rogers, a postdoctoral researcher at the Friedrich Miescher Lab of the Max Planck Society.

Zebrafish (Danio rerio) have been used since the 1930’s in a range of biological studies, including investigations into environmental pollutants and health, embryo growth, brain function, and disease development. Why have zebrafish become such a popular model organism?

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Topics: Genome Editing

Mouse Modeling, Part 2: Breeding and Crossing Mice

Posted by Aliyah Weinstein on Aug 6, 2019 8:55:01 AM

In Part 1 of our mouse modeling blog series, we covered techniques that can be used to introduce genetic modifications into mouse embryos. But once you generate a growing colony of genetically engineered mice, what can you do? In this post, we’ll cover why and how to cross mice to create double knockout lines and Cre-lox lines, and how to properly control for genetically engineered mice in your experiment.

As you’ve learned in Part 1, there are many types of genetically engineered mice: transgenic mice, knockin and knockout mice, and conditional knockin or knockout mice. While these techniques are each useful for introducing one modification into the mouse genome, they are not commonly used to introduce multiple mutations. This is because as more mutations are introduced into a single embryo, the likelihood that a mouse will end up with the intended genotype at every allele decreases.

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Topics: Genome Editing, Cre-lox

Mouse Modeling, Part 1: Genetically Engineered Mice

Posted by Aliyah Weinstein on Jul 11, 2019 9:26:08 AM

Mice are a common model organism used to understand mammalian traits and genetically engineered mouse models provide researchers with useful and adaptable tools to perform basic and preclinical research. For scientists new to using mouse models, the possibilities may seem endless - and overwhelming.

In the first blog post in this series, I’ll highlight terminology you should be familiar with before working with mouse models, several common techniques used to create engineered mouse models at embryonic stages, and the pros and cons of different genome editing techniques.

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Topics: Genome Editing

Screening for Successful Genome Editing with Digital PCR

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

This post was contributed by Scott Findlay, a Postdoctoral Fellow at the University of Alberta.

If you’re like many researchers these days, you are ready to take (if you haven’t already) the plunge into the world of precision genome editing. When it comes time to (hopefully) validate successful mutation of your favourite gene, there are several different methods available. Thankfully, there are many great resources available to help guide you through the rough waters of mutation validation, such as this "CRISPR 101" post. Next-generation sequencing technologies are the gold standard but they remain cost-prohibitive for many labs, and are often impractical for small projects. Most researchers instead turn to so-called “mismatch nuclease” assays (e.g. Surveyor® or T7E1) for mutation detection. While these methods paved the way for mutation validation, we found these assays frustrating to work with, time consuming, and minimally informative. In this blog post, we’ll introduce digital PCR as an emerging validation technology. Digital PCR has several advantages over mismatch nuclease assays that will be elaborated below

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Topics: Genome Editing, Plasmids

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