... the NIH, started off with the first keynote presentation. He talked about the impact of the Human Genome...seeing tRNAs because they contain “hard stop” mutations. New techniques used by the Lowe lab have allowed...species for genome engineering. The final keynote presentation was by Nobel Laureate Thomas Steitz of Yale....

...difficult person: 1) Understand your supervisor’s expectations Every supervisor has their own management style...frequently. The easiest way to meet your supervisor’s expectations is to ask upfront: “How frequently would you... details of your work. Describe the problem by stating the facts and explain how it is interfering with...

...genetically modified organisms, and GMOs have a bad reputation. The public is can be fearful of this technology...ended up applying to a business accelerator freely stating that we had an idea, knew how to build it, but ...in 2004, she accepted a commission in the United States Air Force. She left active duty to pursue a degree...

... clearly As a manager you need to clearly state expectations and goals. Yours and your team’s success ... schedule a meeting to discuss the work, set expectations, answer any questions, and (most importantly...

... of many cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently...of normal protein function, and ultimately misinterpretation of the protein’s cellular role. One way to...However, CRISPR protein tagging does have its limitations, especially when trying to perform high-throughput...

...like adenovirus (Ad) or herpes simplex virus to facilitate efficient replication. The most common production...immunosorbent assay (ELISA) (Grimm et al., 1999), quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet...what Addgene uses, check out our ddPCR for AAV quantitation blog post.  We generally do not recommend using...

...our requestors. However, we can again see quick adaptation and steady use of all types of libraries, once...categorized by year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized...total of 23,006 times, including 14,677 unique citations.  CRISPR is a story that centers around the impact...

...proteins that can be easily modified in order to facilitate its entry into the target cells (Beatty et al...tissues with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing... (2016) Viral vectors for gene therapy: Current state and clinical perspectives. Biochemistry Moscow 81...

...this process accounts for a percentage of the mutations seen with TALENs and CRISPR. MMEJ is active during...using much smaller regions of homology, which facilitates easier cloning. An overview of PITCh. ...possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted gene knock-in ...

...Cellular Structures Construct Fluorophore Excitation/emission maxima Tethering partner Subcellular...genes from organisms with high CpG dinucleotide representation. The specific restriction enzymes used in various... 3. The goal of high throughput cloning is to facilitate the construction of plasmids with many components...

...which contains multiple point mutations in the nuclease domain. These mutations result in an inactive RNA-...hits in genetic screens, they do have several limitations. CRISPRko results in a complete loss of function...

...nothing’s ever perfect, neither is AAV. Its main limitation is the small size of its DNA cargo - 4.8 kb at...strategies are being optimized to overcome this limitation, with limited success so far. Luckily for patients...an ever-growing number of clinical trials for devastating genetic diseases such as hemophilia, muscular...

...attend? What is the expectation on level of critique and detail? Who is/are the facilitator(s) of the journal...papers could be more important.  Any slides, presentations, recordings, and other materials should be made...

...protein synthesis. The kan resistance gene, NPTII, facilitates phosphorylation of kan, which inactivates it....when not 100% inhibited, meaning it will induce mutations in the host/plasmid DNA during selection if it...good choice for experiments in which off-target mutations are undesirable. Pros: Works across many species...

...producer cells where there is E1 and E3 gene complementation. In target cells that are infected by the vector...persistent transgene expression in certain cell types (Tatsis et al., 2007), potentially because some of the ...doi.org/10.1038/s41586-023-05949-1. PMID: 36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval, A., Harris-McCoy...

...the new protocols.io beta platform for sharing, annotating, and storing life science protocols. Since we...simultaneously allowing scientists to actively comment, annotate, and edit protocols. After my introduction to ...lab-use tablet? The software also makes it easy to annotate protocols, make them public for others to use,...

...acceptor FRET know-how with aptamers, aptabeacons (or aptatags) were born. Aptabeacons can be created in many...general premise is that an aptamer in its unbound state contains a fluorophore and a quencher in close proximity...brighter version of the aptamer, but it still had limitations in vivo. Using a combination of SELEX and fluorescence-activated...

.... In vivo, these recombination reactions are facilitated by the recombination of attachment sites from...attL-entry-TOPO vector. TOPO cloning adds short end(s) to facilitate cloning into an attL-containing entry vector.... to 4 DNA fragments, in a specific order and orientation, in one tube, into one Gateway vector to produce...

...inflammation, which can cause mesothelial cells to mutate and ultimately form cancerous tumors.  This process...For example, multiple studies show that certain mutations of the BAP1 gene play a role in susceptibility...individuals, asbestos has not been banned in the United States and many other countries, and it is still imported...

...at the same time increasing interdisciplinary citations in their own work. Of course not all research ...ImpactStory, a website that helps researchers go beyond citation rates to measure the impact of their contribution...Archambault. "The decline in the concentration of citations, 1900–2007." Journal of the American Society for...

...get there: I had to send the company detailed documentation of my results, answer loads of questions, and...differences in customers’ personalities and their expectations. If you are willing to listen to a customer’...support roles. Examples are Technical Service Representative, Technical Support Specialist, Technical Support...

...editors, PE6a–g, that overcome some current prime limitations and enable impressive editing efficiencies in...similar or better efficiency than PEmax, the current state-of-the-art. PE6b was very efficient for short, unstructured...transmission, and plasticity and is a popular marker of excitatory synapses in assays like IHC. In fact, Anti-PSD...

...maintaining their friendships. I put “neutral” in quotations here because I would not have considered myself...handy: Empathy: If you often find yourself in confrontational situations with others because you simply cannot...competing, or accommodating, if you find yourself gravitating most frequently towards these approaches, you...

...training and experience, better align trainee expectations with job outcomes, and increase diversity in...need to know in order to even assess the current state of research became more clear throughout the various...information publicly available may realign trainee expectations with actual job outcomes and/or disincentivize...

...research is dependent on a pathology microtome/cryostat, then making a careful decision on the condition...for suggestions to meet each others’ needs and limitations. Everything is negotiable. Try asking for discounts...research, then it’s worth working within their limitations. 3. Always provide value to others This is a ...

...have intricacies that aren’t always mentioned in static written protocols. It is therefore a bit ridiculous...experiments with too few replicates leading to suspect statistical significance (you can read more about this problem...experimental design practices including talking to statisticians when necessary and encouraging interlab peer...

...across the genome) has been implicated in disease states, and targeting the epigenome may make certain processes... spiroplasma (M. Sss1) (termed MQ1.) The Q147L mutation improves methylation kinetics such that cytosine...transduction. Lysine-specific Demethylase 1 (LSD1) Tatjana Sauka-Spengler's lab has deposited pX330a dCas9...

...galactose3 AAV receptor2 Laminin receptor (LR)3 Putative integrin3 1Wu, Miller, et al., 20062Pillay...for mice in vivo. To target... Try using... Citation CNS AAV1, AAV2, AAV5, AAV8, AAV9, AAVrg Aschauer.... (2006). Α2,3 and α2,6 N-Linked Sialic Acids Facilitate Efficient Binding and Transduction by Adeno-Associated...

...of hands-on lab work for a dissertation. I chose to carry out my dissertation research in a lab in California...outcome for a specific combination of genetic mutations. This is a particular challenge in pancreatic ...

...Location, location, location! What do buying real estate and using AAVs have in common? The three most important... location, location! Just like this common real estate mantra, the choice of serotype (which determines...identified with AAV2 (5), AAV6 (6) and AAV5 at excitatory connections (3). These facts beg the question...

...biochemist, a geneticist, a microscopist, a biostatistician, and that one golden-handed technician who ...Andrew, the team’s founder, to create a mission statement in the first few meetings and we’ve volunteered...like kudzu. Think of a soccer player who gets a reputation for taking risky shots at the goal rather than...

...Kishor Jadhav. Mycoplasmas can remain in the dry state for a longer time. When they come in contact with... is required in this process because result interpretation might be difficult. The low density of mycoplasmas...by these bacteria can hinder correct result interpretation. Also, the signals from the nuclear area make...

... most often consist of an enzyme that has been mutated to block its catalytic activity and instead become... later work showed that introducing a Leu28Cys mutation in eDHFR does lead to covalent binding of the ...work in certain applications — the most common limitation is that certain probes are not very good at crossing...

... to assemble several DNA parts in a specific orientation using Type IIS restriction enzymes (Weber et ... researchers to expand the Fragmid toolkit to facilitate other lentivirus- and AAV-based studies. Unified...acssynbio.3c00423. Pérez-González, Ana, et al. “Adaptation of the GoldenBraid Modular Cloning System and...

...previously-designed tyr gRNA, which causes hypopigmentation, thus marking cells that have undergone genomic...recombineering, with 96-99% efficiency for point mutations and 66% efficiency for small insertions. Multiplexing...degradation tag (LVA tag) to achieve specific point mutations in E. coli. gRNAs are delivered on a different...

...Although it was a huge innovation, there was a limitation to this approach. CBEs and ABEs are base transition...repair (BER), the cellular process that catches mutated U’s in DNA and ideally converts them back to Cs... Cas9s that recognize different PAMs. Their computational tool for predicting CGBE editing outcomes and...

...sequence cells to verify excision didn’t introduce mutations Adenovirus Non-integrating Low Medium Generates...and/or oncogenes, potentially leading to the presentation of a cancer-like phenotype in infected cells...stringency test for mouse iPSCs, tetraploid complementation, involves injecting tetraploid blastocysts ...

... had the lowest background activity in the dark state and highest fold induction at 31X. By using a slit...by blue light (470nm), paCas9-1 induced indel mutations via NHEJ (frequency of 20.5%) and induced modifications...significantly alter the system’s efficiency at generating mutations when activated with light and lowered background...

...currently at 4,651 as of April 8, 2021) Find these stats and more on the addgene.org homepage! You can track...of plasmids right below the search bar and more stats further down the page....

...Hub in this blog post. Alongside our efforts to facilitate the sharing of reagents and data, we’re also ... first paper came out last year on sucrose supplementation to increase AAV2 yields and we’re eager to ...

...Breeding, Genetics, and Biotechnology at Michigan State University with the backing of a prestigious Fulbright... : Kingdom's biotechnology research at Michigan State University 9:36-10:29 : Kingdom's transition back...

... community (check out the infographic below for stats on our plasmid deposits and requests). Over the ... the lookout for new ways to help researchers facilitate and accelerate scientific discovery. How can ...

...student? 4:50 - 8:00 : How did you approach doing rotations? 8:01 - 11:25 : How did you decide what lab to...50 : What is the number one question you think rotation students should ask senior graduate students? ...

...The recent sweeping cuts to science funding, limitations to facilities and indirect costs, and increased...scientific research ecosystem, not just in the United States but around the world. As an organization that directly...

...Addgenies! Our team recently grew big enough to necessitate an office move. We've got ~50 Addgenies working...with institutions and tech transfer offices that facilitate MTAs. The great feedback from scientists who ...

...important to consider your experiment.  It will dictate the amount of DNA you need, and at which level ...DNA that is required and, as a consequence, also dictate the required sizes of the bacterial cultures and...

...Steering Committee and working groups with representatives from all areas of the scientific training and...Sciences (NIGMS) will support the development and implementation of collections of evidence-based practices....

...then you'll find the plasmids listed below the citation. An example of these "Associated Plasmids" is ...increasing your work's visibility Help us improve our citations lists We are still growing our "Articles Citing...

... in some form? There is a lot of talk about the state of scientist training in the US and around the world...postdoc-driven biotechnology clubs such as Penn State, Berkeley and Harvard. Students also partner with...

...the Pacific Ocean, off of the coast of Washington state. Using these samples, he was able to isolate two... protein’s structure and also discovered many mutations that alter and improve its function and shift ...