We narrowed to 605 results for: RAN-1

...-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.   Figure 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 ...

...incorporated at the termini of the crRNA molecule (usually 1-3 of the first 3 bps are modified) for optimal stability... 3’ end. This repeat region associates with the trans-activating crRNA (tracrRNA) which interacts with... important cellular RNA molecules with post translational modifications. One of the most common of these...targeting experiments allows you to visualize transfection efficiency and even track localization in cellular...

...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...

... trafficking of these biosensors.   Figure 1: Schematic of iGluSnFR3 (left). GluSnFR3 in pMinDisplay... from Abhi Aggarwal.   Brandán Pedre Pérez: Redox biosensors Brandán Pedre Pérez is a postdoc at... enable multiplexed imaging of neurotransmitter, calcium, and membrane voltage,” says Tian. Figure...Congratulations to the five winners! Abhi Aggarwal Brandán Pedre Pérez Elizabeth Hubbard Haoyi Qiu He Tian...hippocampal cultured neurons, imaged 14 days post-transfection, excited at 505 nm (right). Scalebar represents...hydropersulfide-propagating enzymes 3-mercaptopyruvate sulfurtransferase (MPST) which catalyzes the desulfuration of...an enzyme-bound hydropersulfide that is then transferred to thiol-containing acceptors. These hydropersulfides...

... Science Find the SEND plasmids    Figure 1: SEND combines an endogenous Gag homolog, cargo RNA...Cas9 fused to the reverse transcriptase (RT), which induces two double-strand breaks and deletes the intervening...They further develop a version of SEND for gene transfer with pseudotyped viral particles, expanding options...enzyme from the Liu lab to target opposite DNA strands and specify a large intervening deletion. Not only...deletions and introduce epitope tags and RNA transcription start sites. The lab further demonstrated that...uses paired pegRNAs targeting complementary DNA strands to program large genomic deletions with or without...PEDAR is capable of correcting larger genomic rearrangements as demonstrated in a mouse model of tyrosinemia...

...for RaPID is an RNA motif of interest. See figure 1 below for details on the key components of RaPID. ...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...

...best tips beyond the basics to get you started. 1) Be visible and open to connections This is a site... 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 ...

...manipulation, there are two main decisions to make: 1. Which modification? and 2. Which viral vector should... 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 ...

...of interest and the metal ions.      Fig. 1: A sample containing a polyhistidine tagged protein...and several contaminating proteins (blue, green, orange) flows through the Ni2+-containing resin. The protein...polyhistidine containing contaminating protein (orange). A low imidazole wash buffer disrupts the weak...polyhistidine-metal ion interaction is not disrupted by denaturants (Hochuli, 1988). In some cases, proteins can ...tag and then refolded by gradual removal of the denaturant. Not all proteins will refold once denatured,...

...antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness because they accidentally...spilling over" into another channel's detection range. Though you can compensate for this when reading...

...sites that were previously unavailable. Figure 1: The PAM sequence that the Cas nuclease recognizes...across a number of different sites. Using a combined random and directed mutagenesis approach, the Anna Cereseto...

...advantage this cycle, consider the advice below:  1. Eligibility check Scrutinize every opportunity prior...increase your competitive edge, they alone, are no guarantee of success. Don’t luck into an internship, prepare...

...the output of the analysis performed in experiment 1. Use camelCase: In camelCase, each subsequent word...like a wet lab experiment I use the text editor TextWrangler to record the following, which are later printed...

...with each protein of interest.    Figure 1: The direct approach (left) versus the indirect approach...in light green; and proteins are represented in orange. Created with BioRender.com.    How do secondary...

... an entirely new affinity reagent.    Fig. 1: A comparison of an antibody (left) to several alternative...bonds, monobodies can be easily expressed in transfected eukaryotic cells, allowing for them to be used...an excellent option for many applications, and frankly, if ain't broke, don't fix it!  That being said...

.... See figure 1 for an example of how changing transfection ratios can affect transfection efficiency. ...containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral elements... structural elements, envelope, and transfer gene are transfected into a packaging cell line, usually .... For some transgenes, transfection optimization simply is not enough to get a sufficient titer; this ...may not translate to the target cell line. To overcome this issue, we recommend using a range of doses...systems such that it no longer requires the viral transactivator, Tat. For more information on the different...requirement for additional DNA can impact the transfection efficiency of the packaging cell line and thereby...

...If you already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise ...reagents can be delivered via transfection, nucleofection, viral transduction or injection as either protein...cell line that can be transfected at high efficiency. Lentiviral transduction Cas9 and gRNA can be present... present in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene...inserted into an AAV transfer vector and used to generate AAV particles. Transient or stable expression...Chemical transfection   Lipid mediated Cationic polymers Calcium phosphate Cells Transformed cell lines... lines Transformed cell lines Transformed cell lines Cargo Nucleic acids or protein Nucleic acids Nucleic...

...patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon methylation underlies...paternal grandmothers early food supply influenced cardiovascular mortality of the female grandchildren." BMC...histone modifications, which can influence the transcription of genes by RNA polymerase II, or by the addition...methylcytosine in CpG islands makes  genes less transcriptionally active. The role of epigenetics in studying...to play a large role in a variety of diseases, ranging from cancer, to diabetes, to congenital disorders...isolated population in Sweden found that paternal grandfathers who had experienced famine before their adolescence...adolescence passed on a heritable trait to their grandsons that decreased their likelihood of death from ...

...basis of many molecular biology tools! Figure 1: Common functions of natural plasmids. Created with...lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation (cellular uptake...? Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection Resources on Addgene.org...environment), transduction (viral infection by bacteriophages), or conjugation (direct transfer from one cell...one popular method scientists use to promote transformation, but it isn’t just for lab specimens — there... evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001) Beneficial...humans only noticed last century! Nevertheless, the transfer of plasmids carrying these genes between bacteria...

... inhibitory synapses in the brain.  Figure 1:Schematic diagram of different FingR constructs and... efficient double stranded templates, rather than the more efficient single-stranded oligodeoxynucleotide...generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent proteins... them to localize to the synapse and minimize extraneous fluorescence from other regions of the cell. ...synaptic maturation pattern of hippocampal adult-born granule cells over time that were not previously possible... al., 2020). Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations...lentiviral CRISPR library that targets human glycosyltransferase genes and other proteins related to cellular...

... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...this surface”….where the complex is docked on the membrane of the endoplasmic reticulum. And graduate students...reported cleavage site on the complementary DNA strand. And this is why we love working with researchers...”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting physical...protein must have in order to scan along the double-stranded DNA, recognize a PAM sequence, allow the complementary...pairing between the guide RNA and the target DNA strand, and then activate two different endonuclease active...structure captures the displaced non-target DNA strand and shows the HNH nuclease domain swung around ...

...article, they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several...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...

... little signal detected along the neurite (Figure 1.) Moreover, the group was able to demonstrate that...expression in E. coli. This operon is under the transcriptional control of a strong constitutive promoter (... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover... protein kinase A (PKA) activity at the plasma membrane in living cells using a construct in which TagRFP-T...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy...