We narrowed to 1,054 results for: STI

...homology-directed repair Homology-directed repair (HDR) stimulated by double strand breaks has been widely used ...application, and the efficacy varies widely — it still almost always results in higher ratios of desired...cost of improved efficiency, although both enzymes still approach or exceed PEmax editing efficiencies for...obtain PEs small enough to deliver with AAVs but still efficient at long and complex edits. The team also...additive improvements. These variants are worth testing if you want to make your prime editing as efficient...this tail is degraded, the truncated pegRNA can still guide Cas9 and compete with full pegRNAs for access...engineered virus-like particles. However, there’s still a long way to go before we see prime editors in ...

...Chapter 1 asks “What is science?” That’s a very big question. Zimmer, a Professor of Chemistry at Connecticut...chapters in this section describe the challenges of trusting science in a “fake news and post-truth era,” the...would be complete without the inclusion of some interesting case studies, and Zimmer provides several. In...throughout this chapter. Now, these observations and suggestions are not so much inaccurate as already somewhat...period of time. Though climate change certainly still is and will continue to be a “critical consequence...current period of rapid flux, the chapter might still feel slightly dated by next week.   A stepping stone...

...fifth repeat Nobel Prize winner in history. Interestingly, he also adds to the disproportionately large..., Provost partnered with colleagues at other institutions to form a support network called the MDH CUREs...MDH-focused CUREs at 50 schools, ranging from R1 institutions to community colleges, and has helped provide... and results with peers and faculty at other institutions. Provost believes these networking experiences...Development. 24(5):127-130. Kimball, C., Powers, K., Dustin, J., Poirier, V., Pellettieri, J. 2020. The exon...purification and characterization of a fusion protein: Histidine tag, malate dehydrogenase, and green fluorescent...

...correctly, there’s no point in crystallizing it or testing its function. Small ubiquitin-related modifier .... Arguably the simplest affinity tag is the polyhistidine (His) tag. Small and unlikely to affect function...the tag size quickly – modifying the length a polyhistidine tract does not greatly alter the size of the...biotinylation signaling peptide (Bio), flanked by hexahistidine motifs (6xHis) MBP Large Protein  40 Solubility...combinations, though frequently those combinations still include at least one element from the original TAP...purpose, and you don’t necessarily want them to stick around after that purpose has been served. At this...

...BFP mutant (BFPH66) contains a histidine at amino acid 66. This histidine is encoded by a ‘CAC’ codon that...consists of a mutated mCherry and a downstream constitutively active eGFP. To create the inactive mCherry...T2A site and expressed in cells. mCherry is constitutively expressed to identify successfully transfected...C-to-T base editing event. This edit changes the histidine to a tyrosine generating a GFP variant (GFPY66...markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing reporter toolbox. ...

...creative, usually saying that they’re not artists, or artistic, and therefore not creative. However, if we think...2019) If you’ve taken the time to do this and are still struggling with idea generation, try this thought...to the same problem. If your lab/company/team is still working towards a culture of encouraging dissent...This is why perfectionists tend to also be procrastinators.  If you fall into this category, get out of...easier to work with than nothing at all - they’re still ideas, after all, and that is the goal of this exercise... paint like Frida Kahlo or you’re stuck drawing stick figures, remember your training as a scientist has...

...we are investigating how the production of cancer therapeutic drugs, vinblastine and vincristine, are regulated...expression method enables us to quickly screen and investigate transcription factor candidates and the regulation...transient methods are usually the first choice for investigating gene function, as these experiments can be done...genes and speeding up research. Certainly, there is still more to transient leaf transformations, but we hope...

... spatiotemporal control over its expression. Constitutive or inducible Cre recombinases can be introduced...coding regions. Typically, each gene isoform has a distinct transcriptional start site. Because CRISPRa can...long-term memory formation. The Bdnf gene possesses 9 distinct transcriptional start sites, all of which yield...singled out two Bdnf isoforms (I and IV) for investigation. Co-transduction of primary rat neurons with...notable exception) necessitates such molecular gymnastics for sensitive applications such as CRISPR screening...CRISPRa? Although cDNA overexpression approaches may still be preferable for individual gene activation in ...

... proactively address questions about your application. These could be logistical, like briefly explaining...and it does take a bit of time to craft one. We still think they're useful, so we've updated our cover...and used a ddPCR assay to answer an experimental question. Now, the hiring manager knows that you can design...most important skills for the position]. I am enthusiastic about [something that you love and is important...any unusual circumstances or proactively answer questions the hiring manager may have. The closing paragraph...

..., a pooled library has hundreds to millions of distinct plasmids, all of which need to be represented ...The size and composition of your library can drastically impact what conditions it needs to properly amplify...pre-made libraries aren’t quite plug-and-play: you’ll still need to titer and optimize before you can transduce...your research. And, of course, if you have any questions, don’t hesitate to email us at [email protected], this is a useful place to ask experimental questions.  McDade, J. R., Waxmonsky, N. C., Swanson, L.... CRISPR libraries. Protocols from the Broad Institute: A variety of protocols and guides regarding pooled...

...echoed in the other testimonies we gathered. The program definitely caused a positive stir and was recognized...invaluable to me as a bench scientist working at an institution where training opportunities weren’t always available...were connected to that Tweet Up in 2013. These existing connections brought in folks I didn’t yet know....  They liked us. They really liked us.  The testimonials from participants in Binning Singletons at ASM...Heather Hendrickson, Jonathan Jacobs, Mo Kaze, Christina Kellogg, Heather Martinez, Pat Schloss, Tara Smith...

...in what they can name a gene and many of the interesting ones come from the fly community. Not surprising...the Shivdasani Lab from the Dana Farber Cancer Institute! Sonic Hedgehog, an important regulator of embryonic...got its name, we need to go back to 1980, when Christine Nusslein-Volhard and Eric Wiechaus first identified...cytokinesis. Why name this protein after a vegetable substitute for spaghetti? I actually couldn’t find the specific...failure of primary spermatocytes to enter meiosis. Testis of cookie monster mutant flies are filled with ...1189. PubMed PMID:1905980. Nüsslein-Volhard, Christiane, and Eric Wieschaus. "Mutations affecting segment...

... neuronal populations in the brain. The Allen Institute and the Janelia Research Campus in the USA have...classic probes. To overcome the limitations of existing FP and epitope tags, researchers in Loren Looger...set-up. In contrast, smFP tags can be recognized by distinct, fluorophore-labeled antibodies with very specific...smFP_FLAG than GFP.   3. Enhanced ability to distinguish weakly expressed and/or similar proteins Through...proteins than standard tags and can be used to distinguish highly similar proteins. For example, N-cadherin...terms of sequence and thus very difficult to distinguish using standard antibodies. By fusing N-cadherin...

...by enhanced green fluorescent protein (EGFP) reconstitution. (a) Scheme of validation for DSB mediated ...mediated EGFP expression cassette reconstitution. When the target sequence was digested by sgRNA guided Cas9 ...SSA, single strand annealing) took place and reconstituted the EGFP expression cassette. (b) pCAG-EGxxFP...details on this plasmid, see Mashiko et al., 2013). Testing the efficacy of your gRNAs is then a simple matter...of gene modification in vivo. Find Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9 gene...require you to house mice that may have no mutation, costing money and space. As a rule, screen all of your ...

...promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent...lentiviral infection, happy virus is the second. When harvesting lentivirus, some labs prefer to collect and pool...the positively charged polycation, polybrene. Interestingly, one study found that alternative polycations...routinely produce lentivirus may want to consider testing a variety of cation and serum sources to ensure... the best conditions for specific applications, stick with them! Changes in such variables as cell density...

...Leong Chew is a Junior Principal Investigator at the Genome Institute of Singapore. His lab focuses on...with Wei Leong Chew, a researcher at the Genome Institute of Singapore who recently started his own lab....work to an independent position at the Genome Institute of Singapore. Chapter 3: Biology research and ... Singapore. Chapter 4: Life as a new primary investigator. Coming up with new project ideas, applying ...

...multiple streams for composting and recycling in the event the object in question cannot be washed or ...reuse, reducing the need for the production of new plastic.  Recycling and reuse is not limited to the office... however. We participate in many styrofoam and plastic recycling programs in the lab as well. For a more...Practices to the Lab.”  Whether participating in an existing process or proposing new programs, we're always...

...advantage for researchers looking to generate long-lasting changes in cells. However, for some applications... genome. Without a working integrase, IDLVs can still deliver their genetic payload to the cell but cannot... IDLVs were first discovered by researchers investigating HIV integrase mutants in the early 1990s (Leavitt...in several patients developing T-cell acute lymphoblastic leukemia due to insertional mutagenesis (Howe...avoid the risk of insertional mutagenesis while still allowing for stable expression of the transgene....of SCID-X1 patients. The Journal of Clinical Investigation, 118(9), 3143–3150. https://doi.org/10.1172/...

...mutations, we still weren’t entirely happy with our editing outcomes and in many cases still didn’t get ...must undergo homology-directed repair (HDR), a distinct cellular DNA repair pathway. Accomplishing this... be screening a total of a few hundred clones - Still better than 10.000, right? (Kwart, Paquet, et al...reads that we studied by deep sequencing, but interestingly (and to our frustration) in many cases we only...currently a Professor for Neurobiology at the Institute for Stroke and Dementia Research, Ludwig-Maximilians-University...

...cells on a flow plot, you might have to do some testing or ask experienced colleagues.   Figure...flow analysis I can spot my lymphocyte population distinct from cellular debris and other larger cells. Note...following gates, cells that are not of interest can still be gated out later. Based on personal preference... many cases a stain will not result in clearly distinct positive and negative populations. Oftentimes,...read an FSC/SSC plot and how to use H vs. A to distinguish single cells from duplicates. Lastly, you learned...Harvard Medical School. His projects include investigating negative regulators of T cells and cytokine ...

... the Institute for Protein Innovation (IPI), a Boston-based, nonprofit protein research institute, partnered...researchers can measure this conformational change to investigate the role of specific integrins in cellular processes...subunit. Using these affinity reagents, the investigators inhibited other RGD-binding integrins, individually...for cell adhesion. The results surprised the investigators, challenging the long-held belief that αVβ3 ...IPI. Caitlin Faulds is a science writer at the Institute for Protein Innovation. She writes about all things...

...cytosine-to-thymine substitutions in target sequences (Banno et al., 2018). These substitutions were limited ...enable C-to-T and A-to-G base substitutions respectively. Substitutions seem most efficient when using... recombineering took place. This system is distinct from those used in eukaryotes in that CRISPR isn...targeted and preferentially depleted from gut. While still far from the clinic, CRISPR-Cas3 systems have been... Sharp JD, Xu R, Prisic S, Husson RN (2016) Investigating essential gene function inMycobacterium tuberculosisusing...

...Even if we stick to citation counts for a moment, it’s clear that articles in less-prestigious journals ...proportion of highly cited articles appearing in lower-prestige journals has risen steadily from 1995 to 2013.... in citations for older publications. (As an interesting aside, Nature recently reported that the most...of another, perhaps it’s worth reflecting on a question of impact that only you can answer: How would ...

...Feng Zhang (of the Broad Institute of MIT and Harvard and the McGovern Institute for Brain Research) – and...I have an important question: How much coffee does it take to fuel more than 30,000 neuroscientists over...Though, to be fair, I think this is a conservative estimate. As a graduate student, I drank 3-4 cups of coffee...Zhang was awarded one of this year’s SfN Young Investigator Awards and we couldn’t wait to congratulate ...

...portable microscope. Less fluorescence admittedly, but still so beautiful! Enabling the scientist in everyone...buoyancy devices” (also known as gas vesicles) as acoustic biomolecule-based dyes. This technology is widely...about ‘Using ultrasound to image bacteria in vivo: acoustic reporter genes’ on our blog. Creative approaches...telling a story makes scientific topics both interesting and approachable!   The grand finale of visual.... References 1. Bourdeau, Raymond W., et al. "Acoustic reporter genes for noninvasive imaging of microorganisms...

... Human Dynamics Laboratory identified five characteristics of very successful teams. Check out Joanne'...if others are involved. They may feel they are wasting their time. The manager may appear to be too busy...to get people sharing.   Frequently ask direct questions for specific feedback in non-public settings. ...demonstrate patience, pause; A = ask at least one question to clarify the situation; C = confirm that you...employees who volunteer constructive feedback and suggestions. Consider offering rewards (gift card, afternoon...

... suffer any less acutely.  The Rare Genomics Institute works to help those who suffer from rare diseases...Through their BeHEARD Award, the Rare Genomics Institute partners with companies to provide technological...Elia Di Schiavi is a Research Scientist at the Institute of Biosciences and Bioresources in Naples, Italy...of spinal cord motor neurons. This disease is estimated to affect anywhere from 1 in 6,000 to 1 in 19,...prioritize which genes they will use to develop diagnostic tools and animals models of the disease in partnership...

...promoter regulating the gene of interest with a constitutive, robust M24 promoter. Originally isolated from...from the Mirabilis mosaic virus, the constitutively active M24 promoter has been modified with duplicated...this new vector, pSiM24, can also be used with biolistic methods, making it useful for engineering plants...large gene transfer. The M24 promoter provides constitutive, high-level gene expression and is functional...antibiotic resistance gene is controlled by a weaker constitutive Nos promoter, and it can be used to select both...

...in December were developed as an answer to that question. Additionally, a concurrent Nature Biotechnology... really not possible before,” said the Broad Institute's Ophir Shalem. “Whole regions of the genome which...delivery vehicle, Neville Sanjana, also of the Broad Institute, said they were able to produce a very diverse... of one of the Science papers, told the Broad Institute. “This will greatly aid efforts to understand ... RNA Library. Koike-Yusa H, Li Y, Tan E-P, Del Castillo Velasco-Herrera M, Yusa K. Nature Biotechnology...

...Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology. If you are reading...culturist has to deal with this problem. It is estimated that mycoplasma is responsible for up to 60% of...always use a second assay to be 100% sure. Before testing for mycoplasma, the cells should be in continuous... for low levels of contaminations to grow. For testing, the medium should not have been changed for at...regularly and dampness of any kind should be avoided. Distilled water provided to maintain the humidity should...Jadhav, MSc is a Research Assistant at the MGMs Institute of Biosciences and Technology in India. Jadhav...

...was in the lab, if you read a paper and saw an interesting construct you wished to use, you could either...we share? NanoLuc® Luciferase-based tools are fantastic examples of technologies that can accelerate your... fusions for imaging Fluorescent proteins are fantastic imaging tools but their use is limited in vivo... activation of other fluorescent probes but can still transfer energy to the fluorescent protein acceptor...create a calcium sensor with the following characteristics: No photoexcitation High Ion specificity High...Yang, J., et al. (2016) Coupling optogenetic stimulation with NanoLuc-based luminescence (BRET) Ca++ sensing...

...called polyhistidine tags, typically include six consecutive His residues (or more). Histidine bonds with...naturally occurring histidine-rich proteins in mammals. Mammalian purification is still possible but requires...cleaved off from the fusion protein while it is still bound to the glutathione resin, eliminating the ...

...Controlling Stimulation Some studies rely on delivering reversible or varied amounts of stimulation to receptors...are: Timing Targeted manipulation Controlling stimulation Invasiveness  Choosing the best set of tools...injected, this is not as precise and the offset of stimulation is gradual. Invasiveness Both optogenetic and...intracranial implant Noninvasive Controlling Stimulation ExogenousEasily controlled EndogenousNot easily...

...recurring meetings, you should share interesting results or ask questions to your collaborators. This helps...person responsible for incorporating everyone’s suggestions into a final version of the document. Set and...to specialized microscopes and allowed me to investigate my protein of interest in a French clinical cohort...’re looking to improve communication within an existing collaboration, or start something new! Additional...

...number of extinct transposon sequences and then testing various combinations of changes. The basic functional...transposase system - one set of vectors for robust constitutive expression (pSBbi derivatives) and one set of...inducible expression (pSBtet derivatives). The constitutive and inducible expression vectors were developed...16 variants with different selection markers (blasticidin, neomycin, puromycin, hygromycin) and fluorescent...robustly express your gene-of-interest, either constitutively or regulated by doxycycline. The Sleeping Beauty...

...explores the choices that went into the design! Still seeing our old logo on the top of this page? You...also needed our branding to support multiple contrasting truths. Addgene is a nonprofit with academic ...forming the letter A. It’s clean and modern, but still approachable. We like the subtle dimensionality ...everything we do. Figure 6: An ecosystem of coexisting materials. Blugene’s new look Addgene’s ...not precisely our purpose. “Science is full of logistical hurdles” is a truer statement of the barriers...

...experimental system comes with fewer ethical and logistical challenges than does working with animals. Because...staff from the ABRC are here to support you! From questions about growing conditions, to guiding students ... are available via e-mail and phone to answer questions and provide support. Growing capacity - ABRC outreach...science learning. Whether visiting classrooms, hosting field trips, conducting teacher workshops, or staffing... staffing a booth at local science festivals, ABRC gets the word out about the importance of plant sciences...

...pGP-AAV-syn-FLEX-jGCaMP7b-WPRE) exhibits the brightest resting fluorescence of the GCaMP7 family of calcium sensors...respond to blue light. We recently released a constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent...next 6 months, pending quality control. For an estimate on availability date or to be notified when a ...exhibits high contrast between peak fluorescence and resting fluorescence. It is useful for activity imaging...exhibits high contrast between peak fluorescence and resting fluorescence.  It is useful for activity imaging...

...people, you will find the answer to your viability question. The miLab you see today is far different than... was viable.  This is not to say that we aren’t still pitching for feedback.  Your venture should continuously...partnerships with universities, companies, hospital institutions and funding bodies. Don't shy away from the ...Although many emails resulted in no response, I was still able to get through to a number of experts, including...make sure you’re able to answer every possible question related to it. First impressions are everything...

...of the population as a whole, scientists gain statistical power in their observations. However, while flow... of FSC versus SSC that allows them to observe distinct cell populations and rule out dead cells and debris...complexity, these cell types will separate into distinct populations on a FSC versus SSC plot (Figure 2...flow cytometry experiment and will appear as a distinct population. After initially gating the lymphocytes...marker) to separate the B cell and T cell into distinct populations (Figure 2). An immunologist who studies...cells from a whole blood sample will separate into distinct populations of monocytes, lymphocytes and granulocytes...