...ve also carved pumpkins (yes, Blugene made an appearance there too) and shared fascinatingly creepy science...algae cell is just the cell wall and cytoplasmic membrane. Vampirovibrio chlorellavorus will not grown in... the snail, the larvae reach the eyestalk and transform them into a pulsating display imitating a moving...grasshoppers and crickets. When it’s ready to transform into an aquatic adult, it forces its insect host...uninfected grasshoppers, including those linked to neurotransmitter activity and geotactic behavior (Biron et ...

...details how to transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via...CRISPR/Cas9, we began to explore the paired double-strand break (DSB) approach at a variety of loci. We were...following paired DSBs is inversions [3]. These rearranged segments may also be informative as to genomic...Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. ...

... origin of replication (which controls the host range and copy number of the plasmid) and typically include...for further study. Promoter Region Drives transcription of the target gene. Vital component for expression...cell types. Primer Binding Site A short single-stranded DNA sequence used as an initiation point for PCR...vector containing the newly inserted gene is transformed into bacterial cells and selectively grown on...biologists to broadly utilize plasmids across a wide range of applications. Some common plasmid types include...

...chemistry. The society currently has 352 members, ranging from undergrad all the way up to professors. Oxford...Oxford currently has around 700 members and hosts a range of scientific meetings for students to attend. It...society provides talks and career support for a range of different scientific disciplines, as well as ... opportunities for society members as well as arranging collaborations and conferences for their students...Synthetic Biology Society also aims to develop a range of talks for both students and the general public...

...how much easier it makes our lives. Plasmid transformation into E. coli is a fairly inefficient process...aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside...Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin...Polymyxin B polypeptide Alters outer membrane permeability Bactericidal 10-100 ug/mL Tetracycline tetracyclin...Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes...

...recombination, one resulting plasmid carries the transgene of interest (aka the viral genome) and the other...resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria containing the... grow! Our 5 Tips for Troubleshooting Viral Transductions will help with the next steps.   References...recombination of plasmid DNA without affecting mRNA translation efficiency or half-life. RNA 25:507–518. https...common lab E. coli strains Find tips for viral transduction  ...

...reprogrammed into iEPs by retroviral expression of transcription factors Foxa1 and Hnf4α, a process that like...reprogramming (CellTagMEF), at the end of the three-day transcription factor reprogramming treatment (CellTagD3), ...determined by at least day 13. However, distinct transcriptional states were seen as early as day 6. All together...MettI7a1 encodes an uncharacterized putative methyltransferase. Adding Mettl7a1 to the standard Foxa1-Hnf4α...tagging two cell populations in a competitive transplant assay in the mouse intestine, demonstrating its...

...sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand DNA incorporation (ssDI). "Easi-CRISPR...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...a DNA repair newbie - diagrams of resection and strand invasion are like a foreign language to me. But...supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "Mechanisms...

...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why...tell you to pick up the pace Constant worry about grants and renewals What is different about non-academia...outsource a task someone else who can do better, no grant writing Job changes can happen regularly (every ...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

...scientific community. Addgene was awarded a Fast Grant to ensure that the scientific community will continue...-2. The resurgence in popularity of the hACE2 transgenic mouse strain, for example, is sustained by the...University of Iowa (McCray et al., 2006), the hACE2 transgenic mice were designed after it was discovered that...Deposits of new reagents, policies from journals and granting agencies requiring materials to be deposited into... implications for clinical manifestations, transmissibility, and laboratory studies of COVID-19: an observational...

...with viruses or transmissible diseases, so we were left with no choice but to quarantine in our houses,...simpler and the volunteers helped with things like translations, bibliographic searches, etc.     Expanding...we call “asymmetric collaborations.” These are transient partnerships where a researcher makes a small ...blog post about remote collaborations Read our Transferable Skills Guide article about external collaborations...

...from green to orange such that non-growing cells are orange, slow growing cells are orange-green, and fast... and in assays for measuring human serotonin transporter activity. Unger also plans to use the serotonin...choice is the Arabidopsis thaliana - Pantoea agglomerans interaction. But the shape and metabolism of ...

...loci. 3. Cleave target DNA resulting in a double-strand break (DSB) Cas9 and its variants have two endonuclease...positions the nuclease domains to cleave opposite strands of the target DNA. Thus, the end result of Cas9...arrays as found in the bacterial genomes are transcribed into pre-crRNAs containing both the spacer region...RNase III, the tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound...target DNA which is E) cleaved leaving a F) double-strand break. A “gRNA” is a researcher-designed hybrid...

...alone. A few years ago when I started my career transition from bench science to software product management...explaining more of her role, which eventually led me to transfer to her team and begin my product management (PM...science backgrounds, most did not. It was my job to translate complex lab operations workflows into a format... help improve their work. Then, I distill and translate that valuable information back to the rest of ...science companies, try to find internships with translatable skills: working with data, working with people...

... different cell types so it is useful in a wide range of applications. This plasmid was the top requested...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing new shRNA sequences...the necessary elements for packaging, reverse transcription, and integration needed for the production of...

...microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available for antibody conjugation... choice for ELISAs.  Biotin Biotin is used in a range of antibody-based assays. Biotin itself is not visible...Endogenous carbohydrates added to antibodies post-translationally. 4) Modifications made to antibodies, such ...commercially conjugated antibodies available, that never guarantees the antibody you need will be conjugated, or ...Stephenson W, et al (2017) Simultaneous epitope and transcriptome measurement in single cells. Nat Methods 14:...

...Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to ..., and how long their signals last all impact transduction of the message. To study signaling pathways,... game of telephone, often the signal must be transmitted by intermediate messengers. Optical control of...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...feedback loops are a common feature of signal transduction pathways, but are often hard to decipher because...

... Also, many people on LinkedIn don’t appreciate random requests without a personal message and they may...trolls. People don’t like to connect with complete strangers as it borders on my first rule of networking--...a useful way to build or use a network. Total strangers can’t really help or refer you. Do not ask people... lot about requesting connections to complete strangers for informational interviews. Many have had success...considered intrusive and it’s still connecting with a stranger so is highly discouraged. You should be using ...

...Ambika, et al. "Transforming training to reflect the workforce." Science Translational Medicine (2015)...better solution would be to reduce the number of entrants into PhD programs to more closely reflect the ...symposium. Eve Marder, professor of Biology at Brandeis University, has recently published an article ...Among other involvements, he helped to establish grants through the Sloan foundation for postdoc-related...

...editing assemblies are sensor-actuators, either transcriptional regulators or chromatin-modifying enzymes. ...cell surface. When the cells interact, GFP is transferred to the receiver cell, and the receiver cell turns...et al., 2021.   The AID system relies on a transport inhibitor response 1 (TIR1) expressed under a ...These libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent.... Check out the new additions.  Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f, now...

...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit from...spatially and temporally target single cells. Now, Francois St-Pierre’s lab developed a new form of FACS-based...-based Dual-fluorescent DSB Repair) is a double-strand break repair assay for mammalian cells. It's based...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced...

... linked to a certain behavior. It is a very old branch of science, dating back several hundred years, ...genes. A popular method uses Cre recombinase on transgenic flox animals to introduce modifications at the...Timing: High expression levels of the viral vector trangenes typically occur two weeks after injection. Therefore... after several weeks which may also depend on transgene or promoter choice. Behaviors you can study using...Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene transfer to the CNS using recombinant adeno-associated ...

...easy — our tech transfer team will communicate with your organization’s technology transfer office or other... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...require that you send your samples to Addgene pre-transformed in this strain. Hazardous If the plasmid...samples to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new...

...looks like the genetic cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate... plasmid). Viral production then proceeds by transfection of all three plasmids into a producer cell line...HEK-293T, production of viral particles, and transduction of a target cell line with those particles. ...Env, Gag, and Pol genes are not encoded in the transfer plasmid, the component that physically integrates...without endangering yourself or others. Happy transducing! Resources Resources on Addgene.org Addgene...

...pattern opens up the possibility of directional gene transfer, analogous to traditional restriction enzyme cloning... cloning. Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells...gene delivery due to their low immunogenicity and range of serotypes allowing preferential infection of ... yet characterized the potential “sticky end” transgene insertion mediated by Cpf1-generated overhangs...Alkhnbashi OS, Costa F, Shah SA, Saunders SJ, Barrangou R, Brouns SJ, Charpentier E, Haft DH, Horvath ...

...sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability of...infected and would have the potential to express the transduced reporter gene. Generating a FLEx switch to control...level in the mouse. 2003 PubMed PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution...Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping. PubMed PMID: 18614669. PubMed Central...adeno-associated viruses for Cre activated and inactivated transgene expression in neurons.  PubMed PMID: 2286602. ...

... 0.07 ml Skill required: advanced Intranasal instillation Intranasal instillation is very similar to its...behind the eye. IV injections are used for a broad range of substances, as it circulate what is injected ...target the lungs. The main differences are that intranasal is easier, less invasive, and only requires a...Anesthesia Trachea 20–23 (tubing) < 0.07 mL Advanced Intranasal Anesthesia Nose n/a < 0.05 mL Beginner Oral...

...generate a classic double-stranded break, while modulators regulate transcription and chimeric efforts make...Countries shown are the top 10 requesting countries. FR=France; KR=South Korea; CH=Switzerland; AU=Australia.  ...Pyhtila et al., 2023). Our thanks to Rodolphe Barrangou for his authorial contributions to, and support...References Pyhtila, B., Kasowitz, S., Leeson, R., & Barrangou, R. (2023). The Expanding Dissemination and Distribution...

... are sensitive to pH. Anything between orange and light orange usually indicates that the cells have grown...culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red)). Blood pH normally lies...too young and should not be used for important transfection experiments right after they’ve been thawed.... spent organizing, an hour is earned (Benjamin Franklin)! It’s good practice to label your plates and/...

... work has always been focused on gene transfer, from transgenes to plants, chloroplast to the nucleus,...to expand the toolkit. We have included a wider range of terminators than are commonly used as we have...well in plants and give similar stability to transcripts as the ones that people use all the time. Now...Plant SynBio Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...

...requires more photons. For example, moving from transient overexpression of protein fusions to gene-edited... lipoic acid ligase, biotin ligase, phosphopantetheinyltransferase) – These enzymes catalyze the covalent... tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine)– Nonnatural amino acids can be ...improved brightness, photostability, and spectral range. Nevertheless, these fluorophores were designed ...analogs without sacrificing their small size and membrane permeability (Figure 2c). These cell-permeable...

...to be translated into Modern Greek to be easily accessible today. Roughly speaking, we translated our software...see these improvements, you are benefited with reassurance your plasmids will not get lost in a freezer ...Another advantage of Scrum is that it provides transparency to everyone involved about the status of each...accelerating research and discovery. We work with transparency, gladly share our knowledge, and love to collaborate...

...PRIME, PASTE relies on single-stranded nicks instead of the double-stranded breaks used in other insertion...target. In addition, when they looked at the transformed E. coli expressing Cas7-11, they found no evidence...broadly similar to shRNA and Cas13 systems in a range of mammalian cell lines. However, in those same ... of landing sites via Cas9-directed reverse transcriptases, followed bylanding site recognition and integration...

...high-impact journals, and the requirement for a high grant success rate. At this point, candidates are highly...requirements (and maybe also the proposed salary range) should help. In smaller companies, technical support... The benefit is that these skills are highly transferable and will help you enter other roles in project...departments such as research and manufacturing, quality assurance, product management, marketing, and sales (to ...drive to help others. These roles offer an easy transition from an academic lab to industry without being...

...methods of ‘horizontal gene transfer’. Resistance can also arise from random mutations in the genetic sequence...didn’t develop, either through random mutation or horizontal gene transfer, means to defend themselves from...bacterium sharing genes with another, an accidental transmission of genes from one bacterium to another through...

...best gRNA Figure 1: Validation of double strand break (DSB) mediated homology dependent repair ...repair (HR, homologous recombination; SSA, single strand annealing) took place and reconstituted the EGFP...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...fluorescence, the better that gRNA is at causing a double stranded break at the target site, allowing the two halves...

...examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place preference...(viral expression, efficiency of laser light transmission to the target neurons, appropriate stimulation...optogenetics is performed in this species. However, transgenic rat technology is improving and may one day reach... reach the sophistication of mouse transgenics. Everything that has been described is intended for the...References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala to nucleus accumbens facilitates...

...supporting datasets must be logically annotated and arranged, as in an actual publication and not require interaction...responsible for storage.  If datasets are too large to transfer to a journal repository, they are likely too large...albeit unintentional, tendency to work against transparency. This opens science to charges of elitism and...opportunities for research as well as increasing transparency. Thank you to our guest blogger! Jim Woodgett...Will Pay for Public Access to Research Data? - by Francine Berman and Vint Cerf (Science, August 2013) PLOS...

...strong transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...rely on exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate ...worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow manipulation of bacteria...Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled by coupled trade-offs between...

...software in order to generate signals in a reasonable range. If colleagues in your group already have experience...experimentally-specific gates. Two-dimensional staining (quadrant gating) For our last example plot, I will show...cells (CD8+) from dendritic cells (CD11b+) using a quadrant gate. Within my “lymphocyte + single cell + live... two stains in relation to each other. Using a quadrant gate, I can divide my cell population into DN,...for each stain respectively). (B) Instead of a quadrant gate, I am free to use multiple individual gates...