Colorful CRISPR technologies are helping researchers visualize the genome and its organization within the nucleus, also called the 4D nucleome. Visualizing specific loci has historically been difficult, as techniques like fluorescent in situ hybridization (FISH) and chromosome capture suffer from low resolution and can’t be used in vivo. Some researchers have used fluorescently tagged DNA-binding proteins to label certain loci, but this approach is not scalable for every locus...unlike CRISPR. Early CRISPR labeling techniques allowed researchers to visualize nearly any single genomic locus, and recent advances have allowed scientists to track multiple genomic loci over time using all the colors of the CRISPRainbow.
High-throughput cloning, in a nutshell, is the systematic combination of different genetic sequences into plasmid DNA. In high throughput cloning techniques, although the specific sequences of the genetic elements may differ (e.g., a set of various mammalian promoters), the same cloning procedure can be used to incorporate each element into the final construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many biological fields. In synthetic biology for example, high-throughput cloning can be used to combine the functions of different genetic elements to generate non-natural tools such as novel biological circuits or sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies, the combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters is a useful application of high-throughput cloning. MXS Chaining is one such technique and has been used to produce complex fluorescent reporter constructs. These fluorescent reporters can be used to detect structure and protein localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu, 2015).
UPDATE (February 1, 2017): THE TRAVEL AWARD IS NOW CLOSED - AWARDEES HAVE BEEN NOTIFIED AND WILL BE FEATURED IN A FUTURE POST
To commemorate their innumerable contributions to the development and use of fluorescent protein tools and their dedication to scientific sharing, Addgene is opening applications for the Michael Davidson and Roger Tsien Commemorative Travel Awards. These $2,000 USD awards will be open to any masters students, PhD students, or postdocs traveling to an academic conference in 2017 who can demonstrate that fluorescent proteins have or will have an impact on their research.
Topics: Fluorescent Proteins
This post was contributed by guest blogger, Luke Lavis, a Group Leader at the Janelia Research Campus, Howard Hughes Medical Institute.
Chemistry is Dead, Long Live Chemistry!
The discovery of green fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer beholden to chemists and (expensive) synthetic fluorophores. Add a dash of DNA with an electrical jolt and cells become perfectly capable of synthesizing fluorophore fusions on their own. Subsequent advances in fluorescent proteins have replicated many of the properties once exclusive to small-molecules: red-shifted spectra, ion sensitivity, photoactivation, etc. These impressive advances lead to an obvious question: In this age of GFP and its ilk, why should cell biologists talk to chemists?
To better highlight the great content contributed by our bloggers each and every month, we've decided to start an "Editor's Choice" series. Each month, I'll summarize the most popular post of the month and point out one or more additional posts that deserve a peek in case you missed them.