...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...things: fun, teamwork, and competition. And our Halloween celebration this Friday will combine all of these...these. Addgenies have been organizing Halloween festivities for years – searching our photo archives I...destroyed to protect the innocent revelers. Our Halloween costume contests have become an annual tradition...colleagues. Success is sweet – as sweet as all that Halloween candy you’ll gorge yourself on in the coming days...Hopefully, I’ve made clear how excited we get for Halloween and how seriously we take costume design. In case...continue to have an amazing turnout for the annual Halloween costume, so I expect some great costumes this ...

...tools, producing lentivirus, can pose certain challenges. Whether choosing a system that is the best fit...will provide an overview of some of the common challenges associated with producing and using lentivirus...make these optimization experiments a bit more challenging. With these considerations in mind, when planning...

...ve had to navigate the challenges involved in plant synthetic biology. Challenge #1: Public perception ...facilitates plant science. Challenge #3: Intellectual property There is another challenge associated with molecular... of well-characterized genetic tools make it challenging to engineer a specific function in these multicellular...fibers. And because working with plants can be challenging, there are a lot of unexplored areas in plant...manner. At Revolution Bio, we are addressing this challenge directly by developing a plant biotechnology product...inspire and open a new conversation about GMOs.  Challenge #2: Technical obstacles to plant synthetic biology... a product without the use of these parts is challenging, and it’s made even more difficult by the fact...

...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...

...another snail, completing its lifecycle. Halloween genes The Halloween genes are a set of cytochrome P450 genes...incorporate science themes into our fun activities and Halloween is no exception. For the past 10+ years, Addgenies...ping pong table. This year, in addition to the Halloween contest, we’ve also carved pumpkins (yes, Blugene...melanogaster (Gilbert, 2004). Mutations in these genes are lethal, associated with defective exoskeleton formations...1983.3 (1983): 813-816. Gilbert, Lawrence I. "Halloween genes encode P450 enzymes that mediate steroid...company culture at Addgene See some of our previous Halloween costume contests ...

...that this basic concept of gRNA design is still challenging for researchers using CRISPR, and that standardizing...paraphrased in the tweet below. The complexity and challenges of homology directed repair (HDR) are echoed ...s a list of blog posts to help you tackle the challenges of DNA repair (with bonus gRNA design resources...Creating knock-in mouse models with CRISPR has been challenging, with protocols using dsDNA templates achieving...

...going virtual for Addgene’s first AAV Data Hub Challenge, now taking place Dec. 6, 2022! Share your AAV...

...I include in my chalk talk?" So what exactly is a chalk talk? On the surface, a chalk talk is quite simple...present a "chalk talk." However, few, if any, faculty job candidates have seen an actual chalk talk. Their...first exposure to a chalk talk is usually their own. This is a problem. The chalk talk is effectively ... hone a chalk talk. Yet, I often hear, "I have an interview next week and it includes a chalk talk. What...steps to chalk talk preparation: Define your research vision. Organize your vision into a chalk talk format...simple. It's you, a dry erase board, some markers (chalk is rare these days), and a room of 5-30 faculty ...format. Practice. Define your research vision A chalk talk could be your plans for your first NIH R01 ...

...portion of the genome. Mendenhall and Richard Myers of HudsonAlpha (where Mendenhall is also an adjunct faculty...As Eric Mendenhall of the University of Alabama in Huntsville explains it, a major goal in his laboratory...Cas9 system for protein tagging. We spoke with Mendenhall and Myers about their new CRISPR tagging system...transcription factors. Addgene: How does the system work? Mendenhall: The system is really quite simple but it offers...advantages does your system have in this context? Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers... Besides being fast and efficient, what other challenges does this system help to solve? Myers: Only a...works on a large scale. The world is too big now. Mendenhall: So many groups have been struggling with this...

...known as HaloTag is actually HaloTag7, and recent specialized variants run up to HaloTag11. HaloTag is now... (Gallagher et al., 2009). HaloTag: Scientists at Promega developed HaloTag in 2008 (Los et al., 2008)...2008), based on a bacterial haloalkane dehalogenase that binds a synthetic chloralkane (CA) ligand. The current...159 Trimpethoprim  (TMP-tag) Few Easy HaloTag 295 Chloralkane (CA) Many Easy CLIP-tag...proteins (often up to 10-fold more). So, labeling a HaloTag or SNAP-tag construct can help you better detect...) Labeling scheme. b) COS-7 cell ER labeled with Halo-Sec61β with SiR-CA (magenta) and outer mitochondrial...protein of interest. For example, the Cohen lab used HaloTag for their “ticker tape” method for intracellular...

...Cases present specific operational or commercial challenges or questions that clients face imminently. Consulting...on analyzing the situation that presented the challenge at hand and help resolve it through well-informed...this realm, there are a few different kinds of challenges that consultants get called upon to tackle, depending...to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...rewarding parts of the job is being confronted with challenges that are crucial for the success of a drug’s ...reasons why client situations might be especially challenging. For instance, the client might need to get a...the next 3, 5, or 10 years. Complicating these challenges is the fact that they occur in the context of...

...Talk." 4. The chalk talk, simple in principle The first question I usually get about chalk talks is, what... whether there is chalk. Sometimes. It can also be a slide deck, markers, etc. Chalk or markers force ... Developing your Chalk Talk These items should be part of the beginning of your chalk talk. These should... This edition has substantial expansions on the chalk talks and Zoom interviews sections, along with new...what is a Chalk Talk? It is a 60-90 min session with department members to discuss your research plans...alternatives for an approach, if asked. The key to a chalk talk is to effectively convey your message. Key ...." This is only the first few minutes of the chalk talk. Realize that it is rare to make it through...

... making it all work – most of the time. #1 Thou shalt find enjoyment in your work If you are happy at ...Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes, work...and with attention to those around you #4 Thou shalt not bear the burden alone You, alone, are not solely...absolutely equally, which means you only have one-half of the guilt. You can handle that, right? Big tip...get enmeshed in a serious relationship. #5 Thou shalt not sweat the small stuff The house can be messy...yourself; you will make progress over time. #6 Thou shalt throw money at the problem If you can, pay someone...to relax and be with friends and family. #7 Thou shalt create community to help Involve family. If you ...

...Nabet et al., Chemical Biology, 2018.    HaloTag The HaloTag was initially developed as an affinity tag...the Halo-tagged protein in question, it is targeted to the proteasome for degradation. The HaloTag is ...straightforward system – HaloTag and its PROTAC are all that is needed to facilitate degradation. HaloTag is also the...visualization of proteins of interest. A fluorescent Halo ligand can be added to media, buffer, etc., which...proteolysis-targeting chimera (PROTAC) was developed for HaloTag. PROTACs are engineered molecules which target ...involves imaging at some stage, then choosing a HaloTag could pay dividends down the road. It is difficult...

...bacteriorhodopsin; a comparative study of strains of Halobacterium halobium differing in pigmentation." Biochemical...Rhodopsin-like protein from the purple membrane of Halobacterium halobium." Nature new biology 233.39 (1971): 149...opsin genes, and include bacteriorhodopsins, halorhodopsins, and channelrhodopsins, the first two found... excitation, whereas yellow light activates halorhodopsins to silence neuronal activity (Figure 1) (Wiegert...of the DA neurons produce behavioral and electroencephalography evidences of arousal in mice previously...release for more information. References Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of...

...while writing each section. Research challenge The Research Challenge motivates the request for application...how you, the expert, will address the research challenge. Now is the time to make bold statements. While...solution is the best approach to the research challenge. Demonstrating expertise in the field and past...how your innovation will address a variety of challenges and impact the field - its encouraged to dream...time budget writer, you may find this section challenging because it’s hard to determine the exact value... differs from publication writing and may be challenging to master at first. Additionally, reach out to...

...funding, bioethics in the age of CRISPR, and the challenges of science becoming increasingly politicized...., “The Professional Scientist,” addresses the challenges of succeeding in science if you’re anything other...ramifications. Readers unfamiliar with academia’s challenges with publication and funding may be dismayed ...The three chapters in this section describe the challenges of trusting science in a “fake news and post-...describes the author’s thoughts on science’s biggest challenges over the coming years. It is arguably the most...the book? It doesn’t need to. The problems and challenges described in The State of Science have not changed...may be the introduction they need to grasp the challenges you face in your career.  Acknowledgements: ...

...the power of lethal mutation co-selection. Only animals rescued for the conditional lethal mutation survive...selection. Knock-in events tend to be rare, and the challenge with any experiment is in efficient recovery of...co-selection approach relying on repair of a conditional-lethal mutation to identify edits with minimal screening...screening effort. Selecting for repair of a lethal mutation should remove much of the unedited “haystack”, ...viable at 15 ºC, yet exhibit complete embryonic lethality at 25 ºC. This method resulted in efficiencies...Engineering of theCaenorhabditis elegansGenome with Lethal Mutation Co-Conversion and Inactivation of NHEJ...

... N713) and C-terminal (residues 714-1368, C714) halves, rendering the Cas9 non-functional when split but... but regaining functionality when the halves are reassociated. By fusing a photoinducible, heterodimerizing...saCas9 protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly ...Nihongaki et al., 2015   CRY2 and CIB1 from A. thaliana dCas9 Activation of gene transcription Polstein...Polstein et al., 2015 LACE CRY2 and CIB1 from A. thaliana dCas9 Activation of gene transcription Nihongaki... al., 2017 Split-CPTS2.0 CRY2 and CIB1 from A. thaliana dCas9 Activation of gene transcription Kim et... et al., 2019 LADL CRY2 and CIB1 from A. thaliana dCas9 Activation of gene transcription. Label genomic...