...-mAbs. It was recognized that an aberrant IgG light chain transcript was expressed by some of the hybridomas...heavy (blue) chains as driven by two CMV promoters (orange). Figure from Andrews, et. al., Elife  A limitation...IF-ICC.) This method was chosen because cells transfected with a plasmid to express the target antigen ...the restriction enzyme BciVI was present in the aberrant light chain sequence, but was predicted to occur...express multiple subclasses of IgG antibody. Broad range secondary antibodies are commonly used in research...

...tRNAs contain internal transcriptional elements that enhance pol III transcription, potentially increasing...base Csy4 recognition sites from a polycistronic transcript. Although this method shortens construct size...coexpression of Csy4 with Cas9 and the polycistronic transcript. As nucleases may have off-target effects and...matching traditional gRNA-expressing constructs, transcript levels of PTG1 and PTG2 were 3 and 31 times higher... of polycistronic gene organization, and its translation to CRISPR/Cas9 shows great potential for multiplexed...

...community. As a result, I have occasionally pondered translating my research into a startup company. However, ...school and turn it into a biotech company. Transitioning an idea from academia to a startup Mike Koeris... biotech startup, Mike explained, his career transitioned away from the bench and became all about finding...stages in the startup". To gain insight into "the random-walk" of a budding startup, I asked Mike to outline...company may begin to prove the technology. This transition takes around 1 – 2 years. The product is the...into it.” Have any other helpful tips to help transition a project from academia to a biotech startup ...

...interaction (PPI) studies in plants that rely on transient transfection systems using Agrobacterium (such as Nicotiana...indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured. Archon1...also showed that Archon1 is functional in a wide range of animal models such as mice, C.elegans, and Zebrafish...al., 2016)). During Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different...plasmids to study PPI using Förster Resonance Energy Transfer (FRET, Heckler et al., 2015) or ratiometric Bimolecular...

...microenvironment. This results in increased expression of transforming growth factor-β signaling, possibly inducing...let’s take a look at exosomes. Proficient in transportation, exosomes are tools of communication and provide... (micro RNA) and other molecules that can be transferred to other cells, inducing epigenetic changes in...in targeted cells. In this case, exosomes may transport miRNA from drug-resistant cells to sensitive ones...enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate and cannot .... "Intercellular nanovesicle‐mediated microRNA transfer: A mechanism of environmental modulation of hepatocellular...

...and IREB2, which post-transcriptionally regulate iron metabolism pathway transcripts. By using RaPID, Ramanathan... include both scrambled RNA motifs and cells transfected with BirA* protein alone (i.e. no RNA bait). ...changes in RC2H1 protein expression and post-transcriptional regulation of SM1v1-containing RNAs could play...often trick host proteins into binding their untranslated regions (UTRs) to aid in the viral life cycle...IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE binding proteins...

...help with transitioning out of academia (because so many scientists are making this transition). Another...have experience and knowledge to help scientists transition out of the academic sphere into a different meaningful...not always as effective in mentoring for this transition. Second, a coach will help you set goals and ...scientific backgrounds. For a stated purpose of transitioning a scientist from academia to industry, you might...in coaching for communication skills. Industry transition is also an area of specialty for Michelle Capes...

...your expected range. For example, calcium in the ER lumen reaches the sub-millimolar range, much higher...are used to monitor calcium concentrations and transients in living cells and organisms. Unlike chemical... the (1) binding of calcium, (2) allosteric rearrangements of the sensor, and (3) subsequent change in...sensors available that will localize at the plasma membrane. GCaMP-X is a nucleus-targeted GCaMP6 with an ...an extra apoCaM-binding motif to prevent aberrant binding of the biosensor to endogenous calmodulin. Multiplexing...

...patients. And, to be honest, I’ve never enjoyed grant-writing. The more I talked to people with industry...only help. Like any major life decision, the transition felt scary. While I knew two people from graduate... to enter academia, it is easier to make that transition from an academic setting—especially if job applications...pages. In academia, however, writing papers and grants is an inescapable and essential part of being able...’re often required to at least attempt to write grants. If you don’t enjoy or excel at writing, an industry...postdoc will free you of the never-ending cycle of grant-writing that is academic life. Of course, because...

...chromoproteins are just now being developed for a broad range of biological inquiries.  What is a chromoprotein... a gene encoding a blue chromoprotein, after transforming them into your cells, you can simply look for... plasmids (meffRed, eforRed, asPink, spisPink, scOrange, fwYellow, amilGFP, amajLime, cjBlue, meffBlue...Image: Murakoshi et al., 2019. ShadowR is an orange light-absorbing chromoprotein that Hideji Murakoshi...imaging microscopy-based Förster resonance energy transfer (FLIM-FRET) in living cells (Figure 4). They created.... K., Tietscher, S., Edlund, A. D., Jamal, S., Wistrand-Yuen, P., … Forster, A. C. (2018). Engineering...

... non-specific endonuclease (Cas9) and a single stranded guide RNA (gRNA). The ~20 nucleotide targeting...of wild-type Cas9 and a gRNA generates a double-strand break in the target DNA, which, when repaired through...scaled up for genome-wide screening due to the broad range of potential target sequences and ease of generating...heterogeneous population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within.... Pooled gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries... you can skip the steps above! Cells are then transduced with lentivirus containing the CRISPR library...

...in a plasmid. During the propagation and/or transformation process, some bacteria will either never receive... is nifty because the recovery period after transformation (the dreaded 60 minutes shaking at 37 degrees... available, cost-effective, timesaving for transformations. Cons: Prone to satellite colony formation ..., interchangeable with Amp, timesaving for transformations. Con: Expensive.   Kanamycin Kanamycin, or...selection gene. However, it does require a slower transformation recovery period than amp or carb (60 minutes...confers mammalian resistance to G418. Con: Slow transformation   Spectinomycin/streptomycin Spectinomycin...

...2 nuclease, Cpf1 (CRISPR from Prevotella and Francisella), was annotated in several genomes and is classified... Cas9’s shortcomings - namely its blunt double stranded cleavage and G-rich PAM requirement. Zetsche et...one to characterize the Cpf1 nucleases. Using Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid...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...exciting both in terms of basic science and translational applications. This discovery of this type V ...

...about? In March 2014, the White House issued a memorandum to the Secretary of Labor to update the FLSA,...postdocs in the final ruling was, for a lot of us, frankly quite surprising. Consortia of higher education...issued, a joint blog post by Director of the NIH Francis Collins and Secretary of Labor Thomas Perez, Fair...exactly how postdoctoral salaries are changing. Transparent salary information for postdoctoral positions... of postdocs at different points on this salary range. We have been able to gather data on changes to ... raising salaries. In addition, both Brown and Brandeis universities have stated that they consider postdocs...

...aims do not need to be grant aims. Each aim could be the basis of a single grant. Either way, think realistically...talk could be your plans for your first NIH R01 grant proposal or a 10-year research plan. I suggest preparing...is a significant need for healthy livers for transplants. Growth of new livers in culture could resolve..., neither of these can substitute for a liver transplant. My lab will combine bioengineered scaffolds ... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

...project to the NAc, where enhanced glutamate transmission leads to locomotion sensitization. (for review... Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC projections to the NAc and decreased...injected Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent IL-1 beta overexpression...the animal behavior. With the availability of transgenic animals and viral vectors, scientists are combining... they injected AAV2 particles into a Drd1-Cre transgenic mouse to deliver Cre-dependent expression of  ...Alpár, Alán, et al. "Hypothalamic CNTF volume transmission shapes cortical noradrenergic excitability upon...

...excellent choice for fusions with integral membrane proteins and membrane associated proteins (i.e. GPI-anchored...these compartments undergo significant post-translational modifications including glycosylation, disulfide... addition of N-glycans [9]. Both of these posttranslational modifications can disrupt FP folding rendering...including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...oxidizing environments when targeted via the Sec translocon." Traffic 12.5 (2011): 543-548. Pubmed PMID: ...

...purification of tagged transcription factors in mammalian cells and transgenic mice.” PNAS 100.13 (2003...and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic acids to produce...lipogenesis, amino acid metabolism and energy transduction. Biotinylation is the general process by which...is responsible for this highly specific post-translational protein modification. Most organisms have less...enzymatic biotinylation of proteins: A post-translational modification of exceptional specificity.” Trends...

...plasmids for use in CRISPR and Tol2 transposase induced transgenesis.  Xenopus Xenopus, or frogs, are one... areas of microbial research like metabolism. Transport, biofilms, and much more. Because of it’s BSL-...have created molecular biology tools to generate transgenic axolotls for an array of different studies. Elly...popular tools for studying frogs include egg transplants and microinjections. Injections of mRNAs, for...Arabidopsis. There are also important tips for plant transformations and the use of Arabidopsis in the classroom...

...size, multimers may exhibit reduced transformation and transfection efficiency compared to the monomeric... Elfinger M, Hartl D, Rudolph C. Transgene expression of transfected supercoiled plasmid DNA concatemers...plasmid multimerization is to avoid growing the transformed bacterial culture into the late stationary phase...the linearized band, ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form...