...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...

... After all I was only going to be interviewed for 1 hour so having a broad enough insight was going to...promotions or further education may allow you to transition into various realms of business.  Thank you ...

...-fDIO mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg...expression, researchers often need to determine these transduction patterns empirically for their own experiments...AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon Harvey 50457 AAV2 pAAV-hSyn-DIO-EGFP Bryan Roth...system takes advantage of the extensive supply of transgenic reporter lines that supply a variety of GFP expression...

...the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two MS2, PP7, or boxB sequences...the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear processes...

...as an endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head ... knockdown efficiency across the 11 candidate transcript targets analyzed. CRISPR-Csm presents an attractive...! Colognori D, Trinidad M, Doudna JA. Precise transcript targeting by CRISPR-Csm complexes. Nat Biotechnol...

...instead conveniently provide them on your smart phone. 1. Lab Archives by LabArchives The single, most important...MSDS for a lab chemical of choice. Now, I can’t guarantee every reagent in your chemical closet is in this...this app. However, if you get quizzed randomly about a hazardous chemical and can’t find your lab’s MSDS...

...present in the data such as synapsin, hsyn synapsin-1, human synapsin, hSynap. Matching We capture multiple...categorization of our information in order to more granularly customize the search engine, and a scoping mechanism...example, matches of a name in the depositor field will rank higher than matches of the same name in the publication...

...Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One of my favorites is making the...work for almost 20 years. It is good to be a bit random because you never know which relationships will...These are all normal outcomes. If you have a broad range of mentors you will always have people you can turn...

...techniques?  Let us know in the comments! References 1. Bacteriophage P1 site-specific recombination. I. ...A double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...maintained).  Translocation: If the sites are on separate DNA molecules, a translocation event is generated... expression – placing a series of transcriptional and translational termination features with loxP sites... pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells, and/or tissue-...that there is virtually no chance of it occurring randomly. Therefore, inserting loxP sites at deliberate...Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination...

...Blood cells are not transduced efficiently with adenovirus and AAV.   Table 1: Types of viruses... Retroviruses are single-stranded RNA viruses that use reverse transcriptase to synthesize complementary...for most experimental needs, and can transduce (infect) a wide range of cell types with a high efficiency... such as a heterologous envelope or a post-transcriptional regulatory element can be used for either type...Adeno-associated virus (AAV) is a small, single-stranded DNA parvovirus. AAV can infect humans and some...response as retroviruses. This virus can also transduce dividing and non-dividing cells, making it conducive...issue. Adenovirus Adenovirus has a linear, double stranded DNA genome which is non-enveloped. It can infect...

...site. Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...the co-transduction by two viruses would lower editing efficiency, the split editors transduced at levels...capacity of 4-5 kb limits the delivery of larger transgenes, such as base editors which have a gene size ...-opsin GEVIs. With an increase in the neuron’s membrane potential, Ace absorbance increases and results...promoters, CRISPRi parts, selectable markers, transformation acceptor vectors, and sequences for genomic...plasmids using the Cyanogate Kit and subsequent transformation into cyanobacteria. Image from Vasudevan et... evaluated CreLite for cell tracing using the transgenic zebrafish model, ubi:zebrabow. In this system...

...and CasPEDIA have got you covered!     Fig. 1 – CasPEDIA Cas ID display information for a given ...Cas enzymes) cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the...cleaves ssDNA in trans. In sum, CasPEDIA documents the target substrate (DNA/RNA and strandedness) along with...Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability...features: CasID CasPEDIA will display both the cis and trans activity of each entry. For example, SpyCas9a (one...can manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.).....32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019). Evolutionary classification...

...her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization and dynamics... has recently published techniques to follow translation in living single cells. However, before CRISPR-Cas9...the Max Planck Institute for Brain Research in Frankfurt am Main, Germany. She presented a novel technology...to be invited to learn from them at the MPI in Frankfurt am Main and perform similar experiments. Visualizing...Both antibodies have a short antibody specific DNA strand added to them. Only if the proteins of interest...are in close proximity to each other can the DNA strands take part in rolling circle DNA amplification. ...microscope she likes to travel, discover new restaurants, go to concerts and bake. you can contact her...

...into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct. Image from Wen ...Wen et al., 2018). SRIRACCHA uses the Piggybac transposon to stably integrate a reporter gene that contains..., you’re ready to edit. These cells are then transfected with several different constructs for the reporter... gRNAs by simply changing the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The...plasmid.  With this set up, all successfully transfected cells will express RFP while cells containing... Lastly, once a mutant is identified, the PB transposase can easily and seamlessly remove the integrated...Knudsen C, Ásgrímsdóttir ES, Rahimi K, Gill KP, Frandsen S, Hvolbøl Buchholdt S, Chen M, Kjems J, Febbraro...

...guide outlines these components.     Fig. 1 - Viral vector components of retrovirus  Adeno-associated...the delivered cargo (often a gene) is called the transfer plasmid while the other plasmids with packaging... or something similar, and is contained on the transfer plasmid. If expression is the goal, a promoter...Pol, the last component, contains the reverse transcriptase and integrase components to convert the cargo...other to allow DNA synthesis of the complementary strand to yield dsDNA. Unlike LTRs, ITRs are perfectly...delivered and expressed in the host cell from the transfer plasmid. Viral packaging and entry The viral packaging...the primary viral polymerase. E4/E1A activate transcription of other relevant viral genes (including the...

...may begin to prove the technology. This transition takes around 1 – 2 years. The product is the measure...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...into it.” Have any other helpful tips to help transition a project from academia to a biotech startup ...

... UK team members before they log off for the day. 1:45 PM--Write final report. Work on a final report ...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...

...non-canonical amino acid at the site of interest (figure 1). While the codon used is often one of the three stop...factors can negatively impact both translation and fidelity of translation, leading to system-wide effects... incorporation of an unnatural amino acid in translation (Noren et al., 1989). In the late nineties, this...non-interacting with) the cells’ own natural translation machinery; these are often incorporated into ...

...medRxiv.org) in the past few years.   Fig. 1: Stand aside, CERN - it's a new age of accelerating...conference poster or talk, or submitting patent or grant applications; to informal settings like lab meetings..., and increase their impact. Open Access and Transparency Another major benefit of preprints is their ... of listing “Submitted” articles in their CV or grant applications. A number of organizations (such as...be prone to biases or errors, with a lack of transparency in most review processes. Preprints and their...

...in Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while... four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting...modify a given gene. CRISPR-tranposons By combining CRISPR editing and transposons, the Sternberg and Zhang...developed CRISPR-transposons. The Sternberg lab's system called INTEGRATE (Insertion of Transposable Elements ... CAST (CRISPR-associated Transposase) both enable gRNA-directed transposition. INTEGRATE consists of four...multi-spacer CRISPR array, CRISPR-transposons can be multiplexed. Transcriptional repression (CRISPRi) in bacteria...proteins involved in the activation of transcription including transcription factors, phage proteins, and RNAP...

...first of only three such databases, EENdb (published 1 Jan 2013, no longer available), is a simple catalogue...follow him on Twitter @CMacPhD.          References  1. Ghorbal, Mehdi, et al. "Genome editing in the human...should we use all tools as some kind of a meta/Frankenstein app? Or perhaps there is a clear winner, is ...enthusiasts, take a look at sgRNAcas9. Top tools for transferring CRISPR technology to a new organism: For obvious...

... genes with a single viral vector.   Fig. 1: Schematic of multiplex gene editing with DAP array..., lengthy genetic payloads and insufficient transcription on the array have limited the scalability and...

...regarding what makes this group of researchers special: 1. Stem cell scientists care deeply about ethics Ethics...that stem cell scientists are so focused on translational research is encouraging. Patients are key to...

...region: IgM, IgD, IgG, IgE, and IgA.    Figure 1: Diagram of an antibody labeled with Fc, Fab, heavy...that helps attack extracellular pathogens.  Transportation of antibodies to places they can’t reach on...region, can be bound by special receptors that transport antibodies through cells and into different body...tears, saliva, mucus) where it protects mucosal membranes. This isotype is most commonly a dimer with a ...

...give these stabilized promoters a try. References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag, and...number during the transition to stationary phase, Segall-Shapiro et al. placed a trans-acting RepA protein...plasmids modified from pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered...

...speed: base editing before Cas lets go Figure 1: David Liu gives a talk at the Cell Symposia: Gene...induces the DNA base change without a DNA double-strand break. But base editors aren’t perfect. They may...References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable base ...

...accessible to practically everyone.     Fig. 1: Open source hardware builds   Since I started...next few years, with the goal of creating more transparent and reproducible science in the long run (Hohlbein...2020) Hongquan Li, Deepak Krishnamurthy, Ethan Li, Pranav Vyas, Nibha Akireddy, Chew Chai, Manu Prakash, ...

...doesn't work for your application.      Fig. 1: Antibody Selection Flow Chart    What does validated...-independent assays such as mRNA data from transcriptomics; independent antibody strategies that compare...validation does not mean that the antibody is guaranteed to work. Antibody-based assays are complex and...

...many of which are available in rAAVs!   Figure 1: Diagram of AAV components      Cargo capacity...therapies and in vivo laboratory experiments ranging from transient gene expression in a particular cell type... Figure 2: DNA rescue and transduction of a conventional single-stranded AAV (ssAAV) and a self-complementary...genome conversion in the transduced cell nucleus, the single-to-double stranded conversion of the DNA goes...structure can readily fold into transcriptionally active double-stranded form through intra-molecular annealing...Adeno-associated virus (AAV) is a single stranded, Parvoviridae DNA virus, packaged in a non-enveloped...hijack host cell machinery, and synthesize a second strand of DNA before expression of their cargo can begin...

...internalization of two different receptors- angiotensin II type 1 receptor and the β2 adrenoceptor. GPCRs in chemogenetics...different ligands, including hormones, neurotransmitters, photons or odorants, to elicit downstream signaling...seven-(pass)-transmembrane domain receptors, due to the seven alpha helices that transverse the cellular...cell membrane. Heterotrimeric G proteins are important molecular switches in signal transduction pathways...fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription factor from the receptor...initiating signaling pathways in inflammation, and neurotransmission. These receptors can be activated by an array...cellular membrane. The receptor recognizes the appropriate ligand to activate the G proteins. It is estimated...

...see how you use them in your research!  References 1.Tang, Jonathan C.Y., et al. “A nanobody-based system... to create a complete transcription factor, activating luciferase transcription. Tang et al. coined this...Connie Cepko’s lab allow researchers to activate transcription or Cre recombinase activity only in the presence...Using nanobodies to create GFP scaffolds for transcriptional activation The project began with a frustration... in their 293 cell culture system. In cells cotransfected with GFP and the GBPs, GFP linked the two GBPs...this system T-DDOG (Transcriptional Devices Dependent On GFP). In testing the specificity of their system... dsRed, mCherry, and TdTomato do not induce transcription. Thus, T-DDOGs can be used in combination with...