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...the ability to cleave the RNA target.  Table 1: Comparison on Common CRISPR Enzymes Name  Enzymatic Domains...Amplified nucleotides are combined with Cas 13a in complex with crRNA targeting the sequence of interest and...multiplexing; expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although... cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found that LwaCas13a ... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

... research compiled in the ELN. Without such references the record of research is incomplete, making it...commercialization office, the CIO’s office, and research computing. The two guiding principles emerging from the ...the record of research captured in the ELN is incomplete. Thus, to support reproducibility an ELN should...select and deposit individual documents or more complex datasets into the desired repository. They also...

...applying to the specific company or lab. Take some time to research the lab or company via their papers and...place to state why you’re applying to the job, company, and/or lab, and how you feel connected to their...apply for [title of the position] at [name of the company/organization]. I am a [adjective that describes... are interested in working for this lab and/or company. Remember to proofread your cover letter, and ...

...brainstorm session is to start with something completely unrelated to the issue and consider how it could...unique solutions to the same problem. If your lab/company/team is still working towards a culture of encouraging...just as valuable! In 2019, Addgene implemented a company-wide New Ideas Survey. Anyone could fill out the...of which, when implemented, impacted the entire company. It is important to note that the person who submitted... is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism When ...

...sgRNA targeting the genomic site of interest. We compared the pCas/pTargetF system with the updated pEcCas...system.   Advantages of the pEcCas/pEcgRNA system Compared with the original system, the pEcCas/pEcgRNA is...strategy for plasmid curing, providing users with a complete editing process. The MUCICAT system While the ...dehydrogenase expression that is 2.6 times higher when compared to the industrial strain. In our metabolic engineering... pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction...

...necessarily mean that you are a future employee of the company. It does, however, offer you an opportunity to ...like to work, get online and take a look at the company or institution’s career page, where you’re likely...openings. A good approach is to do this first for companies at which you’ve already made a personal connection...result in a recruiter giving you a call. Many companies enlist the assistance of recruiters for identifying...development center, if one exists. Get online! Review company and institution career pages. Make yourself visible...

...fields in order to provide useful solutions to complex challenges. As a cross-disciplinary researcher,...Many of the challenges in science today are too complex for any single discipline to solve them on their... disciplines leading the way, the future of complementing published research with video overviews was ...may never access online journals or even use a computer.  Currently, two of the more popular biology videos...derived from research. The platform uses video to complement traditional print journals and expand our ability...

...in the targeted brain region after stimulation compared to controls without Chr2. c-Fos is a reliable ... to proceed with your behavioral experiments. Comparison of c-fos amongst animals that receive different...Inhibition of neuronal activation is a little more complicated since basal neuronal activity in your brain region...writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow...applying optogenetic tools derived from direct comparative analysis of microbial opsins." Nature methods...

...revolutionised the way we apprehend molecular complexes and is still a very useful tool.    Like a guitar... These issues can be overcome by choosing a complementary pair of fluorophores. To maximize the FRET signal...to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian...Clover pGWF1 Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between...especially when you investigate unknown molecular complexes. But it is a parameter that you have to keep in...

... of the policy comprised of numerous objections and concerns. Perhaps least compelling of these were those...to individual researchers. Yet accurate and comprehensible organization of data is essential for efficient...numbers of figures such that many have multiple, complex panels. PLOS, itself, has not finalized its definition...collections and interpretation of data, publishing complete data, rather than just a single projection of ...

...such as cancer since mice better recapitulate the complex interactions between cancer cells, therapeutic ...Scientists can make a new line of flies in only 6 weeks compared to months to make a transgenic mouse line. Over...be used to easily screen multiple therapeutic compounds in one experiment. Zebrafish (Danio rerio) Since...embryonic development studies as their embryos are completely transparent. Thus scientists can easily observe...model organisms, the C.elegans genome is also completely sequenced and over 60% of human genes have an...

...size, while SSC values depend on the structural complexity inside the cell or on its surface. Bear in mind...on the cell’s size, morphology, and structural complexity.   Height, Width, and Area Within FSC and...each coming with a different size and structural complexity. After several steps of tissue processing prior... for a single cell gate. However, make sure to compare H and A (or W and A).   Live/Dead Gating Having...cells stain positively for both markers. When comparing two markers simultaneously, we commonly describe...

...for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need a Nature/Cell/Science... research is about and what you think you can accomplish. No matter how many times I say this, candidates...specific questions and experiments. Nobody EVER complains that a seminar was too easy to understand. Make...detail. Finally, state how successfully completing the research will advance the field, create a...yourself to me by email or in person. If you feel compelled to repay me in some way, I encourage you to help...

...tough. Is the content accurate? Are we conveying complex ideas in the easiest way possible? Are the shots...Jennifer. I had finally found my footing with the company. I’d identified the ways to get the most out the...things like lighting, camera angle, and shot composition.  So back to the drawing board.  Cycle 3: A Return... and availability, a traditional solution was completely out of reach for Addgene.  So, onto research!...  Figure 4: The remote directing setup, complete with wireless video receiver, camera control, ...

...sword Microtubules are one of the most important components of the cytoskeleton. They provide cells with ...you’re interested in that other most-important component of the cytoskeleton – actin? Actin dynamics control...manipulating Rho GTPases based on BcLOV4. These single-component systems take advantage of BcLOV4’s light-induced...powerful enough, what about altering the chemical composition of the membrane itself? The Baskin Lab previously...K. H., & Chow, B. Y. (2022). Designing Single-Component Optogenetic Membrane Recruitment Systems: The ...

...lots of hustle and bustle, but the lab was very competitive and not very communal. That didn’t suit my collaborative...choosing a training lab, I strongly suggest you complete a work rotation in the lab if at all possible.... of effort, intimidating use of discipline or competence procedures Threats to personal standing – including...atmosphere makes you successful. Collaborative Competitive Papers have multiple authors One author plus...choose a grad school or postdoc lab. Having a comparison can be very helpful in uncovering strengths and...

...programs. The software has two components - a database of pre-compiled register designs and a search function...sites, promoters, genes, terminators, etc) are composed. Fear not – the state machine design space is ...circuits in general would benefit from a more comprehensive characterization of the recombinases at our ...the Lu Lab at MIT. He is broadly interested in biocomputing and biological circuits.     References 1. N...

..., in vitro screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment... cells, an important immune cell type, in more complex and physiological microenvironments. These screens...immunogenic, a necessity for in vivo studies; (3) is compatible with single gene KO and pooled screening approaches...facilitate the isolation of modified cells; and (5) is compatible with the introduction of unique molecular identifiers...

.... Nonetheless, Addgene’s annual Deck the Lab competition drew upon the creativity of the community, who...place-winning entry doubled as their lab’s decoration competition, and each lab member used an impressive variety... started last year: The Christmas Decoration Competition! #ScienceFun #ZoomParty @Addgene pic.twitter....bacteria, fungi, and viruses. Final tree with the complete #MicrobeAdventCalendar ornament set (plus more...

...General Counsel at Atalanta, an early-stage biotech company that is developing RNAi therapeutics to treat neurodegenerative...law. He said, “you know, the patent lawyer at my company seems to really like what she does.” I thought ...patent counsel and ran a group at two fairly large companies, Genzyme and Biogen. They’re very different jobs...corporate work, and negotiating contracts with other companies. It's one of those "every day is something different...

... a good story (1). From a deep sea shrimp to a compact powerhouse of bioluminescence, NLuc is 100-fold...yield, brightness and improved maturation time compared to similar orange fluorescent proteins. In their...more than 2-fold brighter signal from Antares as compared to Fluc in this model. The construct for Antares...transfer of energy from one fluorescent protein to a compatible fluorescent protein. FRET can be measured by ...fusion protein called CalFlux VNT. CalFlux VNT is composed of Venus fluorescent protein, the Troponin C Ca...

...Beth Kenkel To complement our RNA Extraction Without A Kit blog post, we've compiled five methods for...methods to do this. 5. Plasmids 101: Plasmid Incompatibility by Leah Schiesow If you're thinking about ...features two mechanisms that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow...

...interviewed fellow Addgenies about their role in the company. One of the things I wanted to do was create content...were a part of the different departments of the company. I asked questions relating to their roles at Addgene...month, I can say with certainty that Addgene is a company that cares - not only in terms of its employees... Addgenies. I could take an active role in the company and not just feel like an additional person each...

...add another layer of complexity to imaging, as they allow one to monitor compartment/protein dynamics or... may not be appropriate for high-pH cellular compartments. Sattarzadeh et al. have shown that GFP can ...Drosophila, and mammalian cells. Photoconverted GFP is compatible with other FPs since its emission wavelength ...

...expansion, scientists will be able to request an AAV composed of an AAV plasmid in the Addgene repository and...by Addgene for legal approvals and technical compatibility with our production process. This process will...the service to be fulfilled and QC assays to be completed and reviewed. The timeline therefore will be as...follows:  Request review: 1–2 days Prep and QC completed: 6–8 weeks Average shipping time: 1–2 days (USA...

...recombination by KD, B2 and B3 resulted in complete or near-complete excision of the stop sequence. In contrast...toxicity, but with the caveat of partial rather than complete recombination. If you’re looking to use site-specific...this system. Gerald Rubin’s lab sought to make complex genome modifications in Drosophila using multiple...

... the clinic and cures. There is no doubt that companies are swarming to see what can be done with this...the insulin signaling cascades in only 4 months compared to a year or greater with older technologies. ... in the room. He offered the great analogy of comparing the CRISPR/Cas9 system to Norton Antivirus. Antivirus...the code of viruses to give information to the computer to fight the known viruses. Bacteria have figured...

...projects spanning from the identification of plant compounds and insecticidal proteins to the development of... reside within living plants and that produce compounds with various antimicrobial activities (1). She... newly identified proteins and enzymes. These compounds will later get tested for their potential agricultural...alternative source of essential omega-3 fatty acids complementing vegetable oil, a somewhat poor natural source...

...question is: how should the gRNAs be oriented in comparison to each other? The gRNAs must target different...different strands to create a DSB, but this can be accomplished with either a PAM-in or PAM-out orientation....preferences using HEK293 cells. They started by comparing D10A and H840A total editing efficiency in PAM-in...donor with 40 bp homology arms. The HDR efficiency compared favorably to using WT Cas9 with the left gRNA ...

...synthetize all compounds needed for growth, or auxotrophy, the inability to synthesize a compound needed to...an essential amino acid), and a plasmid that complements the auxotrophy (i.e., synthesizes the critical...yeast prototrophy kit containing 23 plasmids to complement auxotrophy. Negative selection: Cells that have...

... had to get used to the workspaces, as well as complete the lab safety and cybersecurity training before...project is a major project given to each intern to complete and present on.    Figure 1: Addgene's...size should be a max of 500 bp (preferably). I compiled a list of three options from the Addgene catalog...Toj-Villagran is a rising senior in high school who just completed their summer internship at Addgene. They plan ...

...Until recently, there were no completely genetic tools that would allow researchers to label just a fraction...Sparse Predictive Activity through Recombinase Competition (SPARC), a genetic tool that allows for the labeling...PhiC31 irreversibly recombines one of the two competing attP sites to drive expression of either a stop...is placed downstream of the rest of the SPARC components and expression is controlled by the upstream ...

... to complete a six month work experience called a co-op to receive a degree. Northeastern compiles a list...Thankfully, my managers set aside time for me to complete an online social media certification on social...petting dogs, and winning the office Just Dance competition.    Special thanks to my manager, Sr. Marketing...

...some extra guidance to understand the specific components that make up a plasmid and why each is important...Drives transcription of the target gene. Vital component for expression vectors: determines which cell ...clone into. Regardless, once the cloning steps are complete, the vector containing the newly inserted gene...used in research or for therapeutics Addgene has compiled various educational resources to facilitate plasmid...

...CRISPR screen data are also available via comprehensive pre-compiled downloads and via a full-featured REST... to stably archived large-scale datasets by computational biologists. Figure 1: The BioGRID Open... from humans and other species. By accurately compiling genome-wide datasets, which are often obscured...

...t guarantee a publication. There is an art to composing manuscripts and the ASM Scientific Writing and...writer, I was unsure if I should possibly take a composition class in the English Department. He suggested... coming to the writing workshop in D.C. had to compose and submit a manuscript that they were working ...portion of the paper will be generated. After you complete a publication quality figure, produce the figure...

...volunteered to do the sampling and I happily agreed to accompany him to YNP the following year. Geoff Schiebinger...microfluidic parallelization to sequence this highly complex hot spring sample. The sample was separated into...cells each for sequencing, allowing for lower complexity with higher throughput and rigorous assembly....ThermoCas9 from the van der Oost lab. Phylogenetic comparison indicates that like GeoCas9 and ThermoCas9, IgnaviCas9...

...and not solely to ensure a profit. Often, these companies believe they possess something that will truly...the public’s perception of head injury risk. In completing a master’s thesis in science communication, I... I had the chance to carry out a comprehensive review of the literature informing the notion of “heading... Department of Science Communication where he completed a master's degree in 2020, his research explores...

...platform that allows us to provide scientists with complete plasmid sequences. After launching our viral service...present SMRT sequencing only works for self-complementary AAVs and the TA-based ligation and tagmentation...molecules in the DNA extract associate with complementary strands, enough double-stranded species exist...adeno-associated virus particles containing complementary polynucleotide chains. J Virol 9(2): 394-396...

...The technique relies on the basic ability of complementary basepairs adenine (A) and thymine (T) to hybridize...deoxyadenosine (A) at the 3' ends of PCR products. The complimentary “T” in the pair comes from a topoisomerase I-linearized...overnight. 4. Transform TOPO Cloning Reaction into Competent Cells: You can use your standard lab protocol ...transforming your reaction into chemically or electro-competent E. coli - excess salt causes arcing during electroporation...