...containing the putative tracrRNA into a plasmid and expressed it in an E. coli bacteria strain. They found that...domain, characteristic of CRISPR-Cas enzymes. In contrast to other Cas enzymes, Cas14 has not been found...that Cas14 could indiscriminately cleave ssDNA in trans like Cas13 and Cas12 do for RNA and dsDNA, respectively...can cleave fluorescent probes of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this...

...complement our RNA Extraction Without A Kit blog post, we've compiled five methods for extracting DNA without...sharing even broader and began working with tech transfer offices to make reagents available to scientists...that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow We added to our...it, wrote these tips for working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection...

...ability to direct enzymes – from nucleases to transcription factors – to specific sequences of DNA has become...and her colleagues led by Zhao described and demonstrated in a report published in The Journal of Biological...Biological Engineering in March is fused to a translational repressor. In principle, however, designer PUF...on a version that she hopes will change the intracellular localization of RNA. “You can think of changing...of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA. J Biol Eng. 2014...

...GFP spreads through the cytosol via cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC...signal. Red signal intensity increases with GFP concentration and saturates around 30 uM. For low-abundance...most useful for labs that already have created a transgenic GFP-containing organism,” says Hanson, “using...unexpectedly photoconvertible GFP proteins to make a new transgenic line could avoid problems in localization that...Green to red photoconversion of GFP for protein tracking in vivo. Sci Rep. 2015 Jul 7;5:11771. doi: 10.1038...

...plasmid construct, most often in special E. coli strains designed for protein expression. Cultures are then...glycosylate proteins and may recapitulate the post-translational modifications added to proteins in other eukaryotes...the MISTIC (membrane-integrating sequence for translation of integral-membrane protein constructs) protein...(2014): e112643. PubMed PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary...Resources on the Addgene Blog Plasmids 101: E. coli strains for protein expression Plasmids 101: Protein tags...

...details how to transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via...genomic deletions We were inspired to produce intrachromosomal deletions based on the experiments of Kim ...CRISPR/Cas9, we began to explore the paired double-strand break (DSB) approach at a variety of loci. We were...Targeted deletions are just one of many available strategies by which biologists can imagine, create, and ...

...life repair genomic damage, specifically double strand breaks. Researchers have long taken advantage of...product is introduced into the cell by direct transformation. A given insert will typically contain both...selectable gene product for isolation of successful transformants. Addgene distributes several ready-to-use, modular...native conditions and allow scientists to not only track the abundance, movement, and localization of individual...markers for combining modifications within a single strain. In addition to the collections featured above,...

...worked closely with the legal partners and Tech Transfer Offices at universities to make COVID-19 research-related...United States, and many universities and Technology Transfer Offices were, at least, temporarily closing, these...scientists, we’ve worked with many scientists, tech transfer offices, research foundations, and funding initiatives...them available to industry, please reach out at techtransfer@addgene.org and we will initiate a conversation...conversation with you and your technology transfer office to discuss the possibilities for expanded sharing. ...

...many applications, the most well-known being transcriptional activation/repression. dCas9 has also previously...could then be used to examine both inter- and intrachromosomal dynamics, and how they change in response ...turned their attention to mapping pairs of intrachromosomal loci. The technique successfully resolved ... this work represents the first mapping of intrachromosomal loci, a major benchmark in characterizing ...for example, the visualization of chromosomal translocations or chromosomal shattering (chromothripsis)....

...scientists and technology transfer offices trust us to work for them with full transparency. We don’t have to ... have been able to continuously improve all the extra services we provide like quality control, curation...members. We also find that, as a nonprofit, we attract partnerships with organizations that share our ...Renee, Shannon, Susanna, Tenisha, Thomas, Tony, Tracy, Tyler, Tyrel, Wogenie, and Xuan Additional Resources...

...vector design. B) Strategy for leak-free transcriptional control. C) Iterative transduction workflow overview...organoids. The Organoid EasyTag System is a straightforward workflow for doing gene targeting, reporter...specificity for SLs in vitro, and they further demonstrated that when tested in tobacco protoplasts, the...

...Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned to industry...breakthroughs often occur when the unexpected infiltrates the realm of focused exploration. Here, we delve...that may benefit your current project. Another strategy could entail immersing yourself in diverse literature...viewpoints influence our encounters. It underscores the contrast between perceiving the world solely through logic...

...image from articles openly available on PubMed Central (also eLife, PLOS and Figshare). Some use cases... research papers as they are not included in traditional publications. By submitting your data to a repository...Trust now accept interim research products (pre-registration, preprints, data, software, micropublications...Currently we can only connect figures from PubMed Central, eLife, PLOS and Figshare but we are working on... on the Addgene Blog Use Apps to Save Time and Track Your Experiments Use video to share your science...

...their goal of PhD training when selecting a research topic. The end point of training is not to become ... the field? What are possible pitfalls in your strategy and how can you mitigate them? If you were to ...become a pro in a field, but to be trained to think and perform like a scientist. Researchers should  pick ...before diving in. Research projects are designed to train graduate students and postdocs to think analytically...

... CRISPR/Cas as a powerful and adaptive defense strategy against life-threatening viruses. Evolving to...too, are famous for their ability to devise new strategies to reproduce within a host cell. A few virus ...Surprisingly, few genes have been identified that neutralize CRISPR/Cas. Earlier this year, expression of ...only deactivate the Type I-F system and are not translatable to the other CRISPR subtypes. Still, the large...well-suited for various innovative biological and translative technologies. Addgene has empowered researchers...

... our values of transparency, reproducibility, and community-building. The transparency begins with sharing...share plasmids with each other. Together, we can transform the antibody sharing landscape as well.     References...Boeckman JX, Manning CF, Nguyen JT, Bechtold H, Dumitras C, Gong B, Nguyen K, van der List D, Murray KD... Improving Reproducibility through Rigor and Transparency of Experimental Methods. Neuron 90(3):434-6....

...Calcium sensors Tracking calcium dynamics that govern such activities as neurotransmitter release is vital...expression, researchers often need to determine these transduction patterns empirically for their own experiments...GP-AAV-CAG-FLEX-jGCaMP7s-WPRE 105714 AAV8 pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter sensors: Dopamine and norepinephrine Neuroscience...research relies on accurate monitoring of neurotransmitters and we are excited to offer AAV for the newest...

...sequences is Proteobacteria - the most frequent transfer of genes is from Proteobacteria to mammalian expression...natural gene sequence is, the less likely it is transferred into another organism in the lab. Predicting ...Christopher A. Voigt also recently published a tool (trained on Addgene plasmid datasets) to predict the lab-of-origin....1 (2018): 4425. PubMed PMID: 30356044. PubMed Central PMCID: PMC6200774.   2. Nielsen, Alec AK, and Christopher....1 (2018): 3135. PubMed PMID: 30087331. PubMed Central PMCID: PMC6081423.    Additional resources on the...

...specialized E. coli strain for protein production and gives a quick rundown of some of the strains you can use...used to generate inversions, deletions, and translocations. It also provides researchers with many different...Juchheim in this post. 3. Plasmids 101: E. coli Strains for Protein Production Addgenie Julian Taylor-Parker...Where Do I Find One? If you need help making a transition into a new job or maybe to an entirely new career...

... different strategies with CRISPR-Cas9. There are so many different variants and strategies out there....Empower and Accelerate Research Discoveries) Award attracted submissions from 99 universities and foundations...desire to understand the early events in the transformation of healthy cells into melanoma cells. They’...diseases at the same time. The limitation of budget restrained us from costly functional analyses.  “We believe...