...inexpensive enzymes. Now, a new toolkit - aptly named GreenGate – offers all of those same advantages to researchers...advanced molecular plant science lab.” Introducing GreenGate As Lohmann and his colleagues describe in the ...ONE paper that introduced the kit in December, GreenGate includes the six most frequently used elements... selection of transgenic plants. The complete GreenGate toolkit, now available at Addgene, also gives ...transgenic plants. In the most practical terms, what GreenGate can do is “save you half a year,” in comparison...Lohmann said. The key component of Golden Gate and GreenGate are type IIS restriction endonucleases. They are... in a single tube. (Additional details on the GreenGate kit and how it works are available here.) Although...

...plate technique. Time for the #BioSci3319 #StreakPlateChallenge part 1. Lab B needs a redo :-( @SandTBioSci... the lab used three different species for the challenge, but one species always outcompeted the other ...it would be a perfect fit for the streak plate challenge. Chromoproteins are a subset of the fluorescent... distinguish colonies from one another in the challenge. To create a successful streak plate, first the...colonies in the last quadrant after incubation. The challenge is usually successful for the students. “They ...colonies. I should probably make it a bit more challenging, potentially by using a thicker bacterial suspension...transformation efficiency.  Aside from the Streak Plate Challenge, Westenberg’s course includes Winogradsky columns...

...built our new search engine atop Elasticsearch, a powerful open-source search engine that can scale to search...users who prefer to use search, our updated search engine is now more powerful, intuitive, and searches across...team identified. The basic function of a search engine is to find the website resources that best match...in order to more granularly customize the search engine, and a scoping mechanism that allows category-specific...category to expose as filters and designed the search engine to return only the filters that are relevant to...information for a given resource, and designed the search engine to weight search results differently according ...more technical details that make our new search engine unique. Most importantly, with this new implementation...

... an engineering school as an undergraduate, I never fooled myself into thinking I was an engineer. The...talk, which focused on the history of genome engineering from ZFNs through TALENs and CRISPR-Cas nucleases...names in the CRISPR-Cas9 field – Jennifer Doudna, Feng Zhang, and Keith Joung. All Addgene depositors! ...huge impact on this field. Advances in genome engineering & CRISPR technology That afternoon's workshop...workshop, entitled "Advances in Genome Engineering", brought us 8 short talks on every conceivable way of examining...systems, modifications to the RNA components, and challenging target genomes, such as the malaria-causing Plasmodium...Addgene depositor, discussed some of the many challenges of creating biological circuits, and I can only...

...).  CRISPR-transposons for bacterial genomes engineering In the Sternberg group’s initial report of VchINT...additions to the growing CRISPR toolbox for genome engineering in bacteria, alongside more established tools...currently studies CRISPR biology and CRISPR genome engineering technologies. Message or follow him on twitter...high-efficiency, multiplexed bacterial genome engineering. Nat Biotechnol. https://doi.org/10.1038/s41587...

...provide a brief introduction as to how you can engineer new transgenic flies to study your favorite gene...about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific Recombination... genome engineering...

...enzymes are the latest tools in the transcriptome engineering toolbox. How were Cas13d enzymes discovered?...property is size - Cas13d enzymes have a median length of ~930aa - making them 17-26% smaller than other... tell how large the genome and transcriptome engineering toolbox will be. It is, however, certain that...References Konermann, Silvana, et al. “Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors...

...method that can be used for cloning or genome engineering and is based on homologous recombination. It ...lambda red bacteriophage and its use as a genetic engineering tool is frequently called recombineering - short...for homologous recombination-mediated genetic engineering.  It can be used to make an assortment of modifications...efficient recombination system for chromosome engineering in Escherichia coli. PNAS 2000 97 (11) 5978-5983...Homologous Recombination-Based Method of Genetic Engineering. Nature protocols. 2009;4(2):206-223. doi:10.1038...and Court, D. L. 2014. Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination. Current...Mammalian Cells CRISPR Methods for Bacterial Genome Engineering Resources on Addgene.org Browse our Microbiology...

...by Benoit Giquel. CRISPR has taken the genome engineering world by storm owing to its ease of use and utility...Waltz, 2016), the potential of CRISPR for genome engineering in plants is also being realized. There are a...plants with the phenotype of interest. Genome engineering, on the other hand, allows for targeted modification...phenotype. In fact, CRISPR has already been used to engineer the genome of many plant species, including commonly...will present a general overview of plant genome engineering using CRISPR, highlight the specific modifications...plants, and outline the various plant genome engineering tools that are available to academic researchers... Addgene. CRISPR components for plant genome engineering CRISPR can be used to knockout, activate or repress...

...students (1, 2), but educators face significant challenges in designing suitable projects. In the O'Connor...Advisors on Science and Technology [PCAST] (2012). Engage to Excel: Producing One Million Additional College...Graduates with Degrees in Science, Technology, Engineering and Mathematics, Washington, DC: US Government...

...career path and why he chose to go into genome engineering. Chapter 1: Early research on CRISPR and what... 6: Projects in Wei Leong Chew's lab. Genome engineering, gene delivery methods, AAVs, and more. Chapter...and gene therapy.   Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.      Additional... genome engineering...

...bacterial CRISPR engineering One common way of accomplishing bacterial genome engineering is done with recombineering...the genus Streptomyces, yet genome engineering remains challenging for most organisms in the genus. Work...discovered in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria...quirks change the way CRISPR-mediated genome engineering functions in bacteria, but have no fear - plasmids...recombineering-based MAGE (Multiplex Automated Genome Engineering) technology (Ronda et al., 2016). pMA7CR_2.0 ...E. coli and T. citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines...create a system for scarless, iterative genome engineering (Jiang et al., 2015). pCas contains Cas9 and ...

...system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan...making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that gap. As Sakuma and...to include even more loci. Multiplex genome engineering demonstrated in human cells They used the Golden...efficient viral vector-mediated multiplex genome engineering. The system now provides an efficient targeting...Sakuma’s protocol. Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector... multiplex genome engineering...

...lab procedure for separating DNA by size (e.g., length in base pairs) for visualization and purification...longer ones. Thus, you can determine the approximate length of a DNA fragment by running it on an agarose gel...ladder (a collection of DNA fragments of known lengths). Last Update: Feb. 20, 2018 Protocol Video Watch... be purifying the DNA for later use, use long-wavelength UV and expose for as little time as possible ...

...absorb different wavelengths of light to varying degrees and most have a specific wavelength that they maximally...based on its absorbance, you need to know the wavelength of light that your substance maximally absorbs...

... (Gibson et al. , 2009). Regardless of fragment length or end compatibility, multiple overlapping DNA ...fragments at a time. Resources and References New England Biolabs (NEB) (Link opens in a new window) sells... window) . Gibson Assembly® is licensed to New England Biolabs by TelesisBio. Gibson DG, Young L, Chuang...

...post-transcriptional regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed...has opened up a world of possibilities in the engineering of complex gene networks. A comparable system...assemble designer PUF domains for RNA specificity engineering. “The RNA binding domain is interesting because...report published in The Journal of Biological Engineering in March is fused to a translational repressor...

...gel electrophoresis, which separates DNA by their length in base pairs. Following electrophoresis, you ...cutting out the bands and you will want to use long-wavelength UV for as short a time as possible to get the...

... DNA polymerase can create overhangs of varying length (typically 10-12 bp) based on a specific sequence...or tag sequences (where appropriate). The primer length is dependent on the T4 Pol "chew back" reaction...

...used for ligation, use only long-wavelength UV light. Short wavelength UV light will increase the chance...Materials Material Vendor and catalog # AgeI New England Biolabs (NEB) #R0552S EcoRI NEB #R0101S NEB buffer...