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CRISPR 101: Epigenetics and Editing the Epigenome

Posted by Mary Gearing on Feb 14, 2017 10:44:08 AM

This post was updated on Nov 29, 2017.

Epigenetic modifications are an additional layer of control over gene expression that go beyond genomic sequence. Dysregulation of the epigenome (the sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting the epigenome may make certain processes, like cellular reprogramming of iPSCs, more efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression, but causality and mechanisms remain unclear. Today, targeted epigenetic modification at specific genomic loci is possible using CRISPR, and Addgene has a number of tools for this purpose!

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Topics: CRISPR, CRISPR 101

Your Top Requested Plasmid in 2016!

Posted by Tyler Ford on Jan 11, 2017 3:47:00 PM

2016 was an exciting year for genome engineering research. A variety of new tools came out including the single base editors, Casilio, CombiGEM, and a variety of pooled libraries. Not all of these technologies were without controversy, and it remains to be seen how popular any one of them will become. One thing is for sure, as is obvious from our most requested plasmid, SpCas9 is still going strong as the basis for many genome editing experiments. So, without further ado, the most request plasmid in 2016 was...

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Topics: Fun, Inside Addgene, CRISPR, pooled libraries

Truncated gRNAs for Regulating Gene Expression

Posted by Guest Blogger on Jan 10, 2017 10:37:46 AM

This post was contributed by guest bloggers Alissa Lance-Byrne and Alex Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering.

CRISPR/Cas9 technology has revolutionized the fields of molecular biology and bioengineering, as it has facilitated the development of a simple and scalable means of making targeted genetic edits. Cas9 is a DNA binding protein that can be directed to virtually any genetic locus when complexed with an appropriately designed small RNA, or guide RNA (gRNA). The gRNA conventionally contains a 20-nucleotide sequence that is complementary to the target site, or protospacer, in the genome. Native Cas9 has two catalytic domains, each of which cleaves one strand of DNA upon binding the protospacer. The resulting double strand break (DSB) stimulates DNA repair mechanisms that can be exploited to either inactivate a gene or introduce a desired genetic alteration.

Listen to Our Podcast Intervew with Alex Chavez

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Topics: CRISPR

With Our New Viral Service, We're Taking CRISPR Further!

Posted by Tyler Ford on Dec 6, 2016 10:30:00 AM

Numbers in the large colored circles are rough approximations of the total number of CRISPR plasmids for that particular organism available at Addgene. Percentages represent the fraction of that total with the indicated function.

One huge reason CRISPR has become such a popular genome editing tool is its developers’ willingness to make their CRISPR technologies available to the academic research community. At Addgene, we’ve helped distribute many of these technologies in plasmid form and are proud to have facilitated their fast adoption. However, in many cases the plasmids themselves are only the starting point for the production of viruses used to deliver CRISPR components to cells or organisms under study. In the past we’ve left the arduous task of virus production to individual labs, but now we’re very excited to provide ready-to-use CRISPR lentiviral preps to researchers across the globe.

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Topics: CRISPR, Viral Vectors

CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

Posted by Joel McDade on Oct 11, 2016 10:30:00 AM

CRISPR has taken the genome engineering world by storm owing to its ease of use and utility in a wide variety of organisms.  While much of current CRISPR research focuses on its potential applications for human medicine (1), the potential of CRISPR for genome engineering in plants is also being realized. There are a variety of reasons to consider using genome editing to change the genetic code of plants, including the development of crops with longer shelf life and the development of disease-resistant crops to increase agricultural yield (2,3). While it is certainly possible to select for desirable traits using traditional plant breeding approaches, these techniques are cumbersome, often requiring several rounds of selection to isolate plants with the phenotype of interest. Genome engineering, on the other hand, allows for targeted modification of known or suspected genes that regulate a desired phenotype.  In fact, CRISPR has already been used to engineer the genome of many plant species, including commonly used model organisms like Arabidopsis and Medicago truncatula and several crop species including potato, corn, tomato, wheat, mushroom, and rice (4). Despite the almost universal functionality of the CRISPR system in most organisms, some plant-specific changes to CRISPR components are necessary to enable genome editing in plant cells.  

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Topics: CRISPR, Plant Biology, CRISPR 101

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