...occur, but good performers move around easily  Table 1: Taking it a Step Further - Big Company v. Small Company...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...

...motion videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time...have the same issues as one doing experiments in France. We can learn with each other through globalized...

...class.   Four ways to improve your next manuscript 1. Be consistent with your figures The figures of a ...worthless. But simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts...

...then scroll on for some wintery delights!    Table 1: New ready-to-use viral vectors in the Addgene repository...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure...

...heterodimerizes with CIBN upon blue light exposure (see Fig 1*). When the group targeted the enhancer region of ...ubiquitin ligase, the Kanemaki lab was able to translate AID to yeast, as well as mammalian and human cell...PubMed PMID: 31026591. Expanding the targeting range of CRISPRCas9 by engineering SpCas9 Article contributed...are also thought to play a key role in the transcriptional regulation of many genes. A number of groups...thus far for these latter experiments, such as transcription factors tethered to dCas9, require long time...

...wide range of input DNA concentrations. This process is explained more fully in the animated Fig. 1 below...plasmids on a plate) will typically have a wide range of input quantities. This variation is problematic...produced from large numbers of samples with a wide range of input concentrations (via proprietary chemistry...differences in input concentration across a 100-fold range (1ng – 100ng) are normalized to two-fold variation...

...produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard features of a ...plasmids from the same group should not be co-transformed, so if you need two plasmids for an experiment... same incompatibility group should not be co-transformed. +Actual copy number varies. See below for ...stocks, agar stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies...

... and can likely be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo...post about multiplexing with Cpf1.   Figure 1: Multiplexing allows researchers to express multiple...humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator)-containing...Multiplexing from a single transcript You can also multiplex gRNAs via a polycistronic transcript. Rather than being...being transcribed from different promoters, the gRNAs are transcribed together and are flanked by specific...simultaneously. Vectors for both transient expression and Agrobacterium-mediated transformation are available. For ...system for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter...

...Article contributed by Susanna Bachle Figure 1: Initial steps in the CRISPR-pooled screening using...Sabrina Büttner’s lab designed a calcineurin transcriptional reporter consisting of a quickly-degraded destabilized...contributed by Jennifer Tsang Figure 4: Double stranded DNA bound by tTALE-FP with the fluorescent protein...

...net negative charge with 4 negatively charged and 1 positively charged amino acids. Myc tags are frequently...some proteins. Protein folding issues, steric hindrances, and destabilization can be brought about by ...DOI: 10.1038/521274a Arribere, J. A., et al., Translation readthrough mitigation. Nature. (2016) 534, 719...

...extension. Now, they split EB1 into two halves (Fig. 1): one containing the microtubule binding domain and...manipulations on cells, and has potential use for a wide range of applications in cellular biology. Van Haren ...fluorescent proteins (FP) with improved tissue penetrance and live cell imaging capabilities. Upon developing...

...defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is actually a map of Joanne ...enough, but I’ll never forget the strawberries. Grand gestures are memorable! Attend events for one organization... want to do: Don’t connect via LinkedIn with strangers. It’s a waste of time. People that don’t know ...

...impartial. The best science happens when scientists have 1) the courage to make mistakes and 2) the integrity...of what I saw are that we, as a society, need to: 1) use more citations for out data so our sources are...I’ve worked tirelessly to get the publications, grant funding, and teaching experience that it takes to...

... lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

...red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system is spread...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...

...individual protein has all the components necessary to: 1. Bind to a guide RNA The guide RNA, which will be ...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...

...through the use of nanoparticles. [6].    References 1. Zhang B. 2021. “CRISPR/Cas gene therapy.” Journal...editing system to correct the cystic fibrosis transmembrane conductor receptor (CFTR) by homologous recombination...Cas9 has been used to generate mutations in a wide range of organisms, but its potential to efficiently correct...Cas-based Gene Therapy in Human Genetic Diseases.” Theranostics. https://doi.org/10.7150/thno.43360 7. Yuxuan...

... will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for...next interview or whether your receive the next grant you apply for. After all, the only value your work...high level to their team.  When you're writing a grant, this means convincing a committee that your research...collaborating with colleagues in other fields or writing grant proposals to be reviewed by interdisciplinary panels...communities gather at ComSciCon's local and specialized franchise workshops throughout the year.  Find workshops...

...into account, the digest pattern depicted in Lane 1 would be expected for a XbaI/NotI digest. The program...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...overlaps with the XbaI cut site (depicted as an orange line) because the restriction site is followed ...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...

...challenges involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the...how they started their company here. The first transgenic plant was engineered over 30 years ago, but plant... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...