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We narrowed to 866 results for: FER

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  1. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    .... While these differences may seem subtle they result in a key physiological difference between lenti-...3rd-generation transfer vectors apart? A3: Addgene defines 2nd- or 3rd-generation transfer vectors based...packaging systems differ between 2nd- and 3rd-generation vectors? What does this mean for transfer vectors? A4...protein and consequently are considered safer. Second-generation transfer plasmids MUST be packaged with a 2nd-generation... Are transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer plasmids...which is rarely more than 30% of the transfer plasmid. Many transfer vectors are SIN. Packaging plasmid...ingredients: the lentiviral packaging vector, a transfer vector including the gene of interest, and the...

  2. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...Mobile-CRISPRi: a modular approach for CRISPRi transfer Mobile-CRISPRi offers two advantages over existing bacterial...chromosomal copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during...disease. However, the rate of transfer was variable. They measured transfer rates by comparing the number... P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates are better suited for...Mobile-CRISPRi plasmids at Addgene! References Auchtung, Jennifer M., et al. "Biology of ICEBs1, an integrative...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria...diverse bacteria species. Existing protocols can transfer CRISPRi into a single bacterial strain, such as...

  3. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...approach is RNA interference (RNAi). The most common method is transfection of short interfering RNA (siRNA...For CD365, they examined two different antibody clones produced by different vendors. The specific epitope... panels, where different manufacturers may have the same clone available with different conjugates. Summary...commercial antibody manufacturers. References and resources References Ayoubi, R., Ryan, J., Biddle, M. ...include a discussion of the Human Leucocyte Differentiation Antigens (HCDM) workshop approach, which focuses...one size fits all. The approaches used will be different depending on the sample and target type. To start... For intracellular targets, we usually test 3 different fix/perms (methanol, PFA-saponin, PFA-triton)....

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ..., A mutant Renilla luciferase, catalyzes the oxidation of its substrate, luciferin, which releases energy... neomycin or luciferase) to create a final lentiviral vector containing up to 5 different elements. Lentiviral...bioluminescence resonance energy transfer (BRET), whereby photons emitted by a Renilla luciferase variant are used to...alternative to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids... issues with fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash...pHlash, that use Bioluminescence Resonance Energy Transfer (BRET) were developed by the Johnson Lab . Characteristics...energy as a photon. The photon energy is transferred to the acceptor fluorophore, a circularly permuted...

  5. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ... directed differentiation To understand how each human TF can contribute to differentiation, the Zhang...atlas. Computational analysis was used to infer differentiation trajectories and identify both up- and down-regulated... scientist at [email protected]! References and Resources  References Joung J. et al., A transcription...patterns determine everything from stem cell differentiation fate to tumor suppression (and most everything...construct. Cells were cultured following a differentiation protocol before performing single-cell RNA ...embryonic stem cells. Transduced stem cells were differentiated and single cell RNA-seq was performed to map...    This atlas linked TF expression to differentiation outcomes as well as global transcriptional ...

  6. Plasmids 101: Codon usage bias

    Type
    Blog Post
    Published
    Sept. 27, 2018, 1:09 p.m.
    ...most organisms on Earth, but different organisms have different preferences for the codons they use to ... organisms have different codon usage biases? The reasons for varied codon preferences among organisms...poorly charged tRNAs, different regions of the protein may be translated at different rates. The ribosome...Organisms often need to express genes at different levels under different conditions. With varied codon usage...constraints have molded which codons are used preferentially in which organisms - organisms have codon usage...your organism of choice. Unfortunately, codon preferences make it so you cannot choose among the possible... evolutionary constraints that lead to these preferences and what can we do about them? Read on to find...

  7. The Golden Stopwatch Award: Recognizing Facilitators of Research

    Type
    Blog Post
    Published
    Sept. 5, 2019, 1:06 p.m.
    ...your TTO have a question? Refer them to Addgene’s Tech Transfer Team: techtransfer@addgene.org Congratulations...Addgene Help Center: Tech Transfer Technology Transfer Information Tech Transfer News: A Reintroduction ...Addgene account for Tech Transfer Offices. They’re packed with tech transfer resources (summaries, electronic...stats and strategies with tech transfer offices. Drop us a line at techtransfer@addgene.org. Albert Einstein...requests from Addgene MTA: material transfer agreement Technology Transfer: the exchange of intellectual property...addgene.org identifies the hold-up. Your Material Transfer Agreement is stuck in “pending.” Fear not, there... to make MTA turnaround faster with technology transfer offices (TTOs). The latest of these efforts is...

  8. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...utilized mice to study how different mutations in leukemia impact different treatment regimens (Zuber ...example, the Stainier lab studied the proliferation and differentiation of β-cells in the the developing zebrafish...β-cells that originate in different regions of the pancreas and produce different levels of insulin (Hesselson...mouse (Mus musculus) Let’s start with the most preferred mammalian model organism of many researchers: ...injecting mice with retroviral vectors that harbored different mutations found in leukemia, the Lowe lab was ... gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces cerevisiae...at a time. Zebrafish embryos are also laid and fertilized externally, allowing scientists to easily manipulate...

  9. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    Published
    March 7, 2023, 2:15 p.m.
    ...test assesses the sum of differences between pairs). Samples arising from different subjects or populations...test assesses the difference of the group means). Unpaired t-tests can also be referred to as two-sample...significant differences between the means of your groups. However, it will only tell you if differences exist...samples are different from each other, or if every permutation of pairs within the five is different. To identify...groups to determine if they are statistically different. They are classic parametric tests and one of ... whether or not two samples are statistically different than one another, a two-tailed test should be ...control group. If you only care if one group is different in a certain direction (i.e., you just want to...

  10. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ..., Addgene uses 'labeling' to refer to antibodies and 'staining' to refer to small molecule dyes. Indirect...first fixed and then permeabilized, using two different buffer solutions. Fixing causes protein cross-linking...nucleus, a different fix/perm kit is required (to be precise, the fixing solution is different while the...plot (Figure 4b). Through this method, differences in proliferation between cell populations can be visualized...divisions).   Figure 4: Proliferation dyes can visualize proliferative capacity and speed of cells...lineage markers, like many of the cluster of differentiation (CD) proteins found on immune cells. However...use 'labeling' and 'staining' interchangeably to refer to any technique that combines a marker with a signal...

  11. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ... virus can be stored at 4 °C and buffer exchanged the next day. Refer to the AAV Iodixanol Gradient Ultracentrifugation... AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid...help mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest ...more details. Pro tips I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple...to grow multiple bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen...restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, like NEB Stable...

  12. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    Published
    Oct. 30, 2018, 1:04 p.m.
    ...maintaining the undifferentiated “stem” state and, Their potency or capability to differentiate into specialized...special types of cells that can develop or “differentiate” into more specialized cells with specific functions...function adequately [1]. Stem cells’ ability to differentiate into multiple cell types makes them useful models... cell has the ability to produce all of the differentiated cells in an organism, including the embryonic...Pluripotency: When a cell has the ability to differentiate into cells of any of the three germ layers: ...detail below [2]. Multipotency: When a cell can differentiate into multiple cell types of the same lineage...name suggests, cells under this category can differentiate into only one cell type. Muscle satellite cells...

  13. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...engineered γ-retroviruses differ from lentiviruses? There are a few important differences between engineered ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...Vectors 101 eBook, 1st edition.  References and resources References Swift S, Lorens J, Achacoso P, Nolan...following components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA ...sequences, which facilitate integration of the transfer plasmid sequences into the host genome. Typically... the host genome upon viral transduction. Most transfer plasmids available from Addgene were derived from...

  14. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...created phages. Transfer of this DNA to another cell is referred to as transduction. Transferred DNA once inside...donor can transfer genetic material to the recipient bacterium. The genetic material transferred is commonly...Horizontal gene transfer (HGT) is the movement of genetic material between organisms. It plays a key ...unique to the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...Various factors promote natural transformation in different bacteria such as growth phase of the cells (Baltrus...plasmid, the genes on the plasmid can give them different phenotypes. For example, this plasmid encodes ...cell. Once inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages ...

  15. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...Tris-EDTA (TE), just Tris buffer, or some other variations. Does it make a difference? Short answer: yes. This...choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose could...TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving the stability of your preparation...can be Tris buffer (with no EDTA) or water. These are two of the choices that companies offer for the elution...preparation can be eluted in water or in your buffer of preference, and it will be stable for years. Plasmid... preparation for a long time. Tris buffer controls the pH, while the EDTA chelates any divalent cations...through their kit. The advantage of using a Tris buffer is that it allows better control of the pH preparation...

  16. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...through horizontal (or “lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation...conjugation (direct transfer from one cell to another). Bacteria need special fertility genes to be capable...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...Here at Addgene, we often refer to plasmids as lab or experimental tools. They certainly are very handy... 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...Historically, colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.” Virulence...

  17. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...host range is determined by the different hosts that a plasmid can transfer to, replicate in, and persist...stable, can be easily cloned in different bacteria, and can be transferred to various experimental hosts... that can replicate in different bacteria and express proteins within different host organisms. pBBR1-...recombinant vectors at Addgene!   References and resources References Doran, K. S., Konieczny, I., & Helinski...host range, identify a few of the features that confer broad host range in plasmids in nature, and provide... they carry among organisms belonging to many different phylogenetic groups (Jain and Srivastava 2013)...be used by scientists across disciplines.  What confers broad host range in plasmids? Not all plasmids ...

  18. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    Published
    Oct. 22, 2019, 12:20 p.m.
    ...your poster Check conference guidelines To begin, you should always follow conference rules when designing...useful to use two different font styles; one for your section titles, and a different one for the body ...distinguished. The UCLA Library offers some Powerpoint based templates with different column numbers for a good... the most of your conference attendance Read all of our blog posts about conferences .... But, before you can show off your work at a conference, you need to first make your poster. While a ...understand that presenting your work in poster format differs in many ways from writing a manuscript. If you ...you have ever attended a scientific conference, or even a lab recruiting session at your university, you...

  19. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...not transfer precisely evenly across the entire blot. So how can you accurately compare different lanes...assuming that all differences in the loading control are due to loading and transfer errors — not to any...reflect massive differences in protein abundance. Validating the assumptions for different loading controls...detection range be linear! Resources and References References Aldridge, G. M., Podrebarac, D. M., Greenough...errors like uneven pipetting or irregular membrane transfer. Figure 1: A loading control is a type ...Researchers use later steps in this chain (blue) to infer or calculate things about earlier ones. In practice...relationship poses problems for proteins of very different abundance, consider the following example gel....

  20. The MTA at Addgene

    Type
    Blog Post
    Published
    July 23, 2024, 2:17 p.m.
    ...There are many different flavors of MTAs, but the Uniform Biological Material Transfer Agreement (UBMTA...process that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer office is...for more information. Happy transferring!    Many thanks to our Tech Transfer team members Maura Coughlin...Addgene, then you’ve been involved in the material transfer agreement (MTA) process. Even though this process...curious about the hows and whys of the material transfer agreement, read on!  What is the MTA?  An MTA...UBMTA) is one of the most commonly used ones for transfers between academic institutions.  What is the UBMTA...the creation of a standardized agreement on the transfer of biological materials in 1995: the UBMTA. Because...

  21. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ... deposited by Promega here.    References and resources   References Allikalt, A., Purkayastha, N., Flad... scientists. Seeing inside these specimens has offered illumination on biochemical processes that are ...Figure created with biorender.com. Fluorescence refers to the emission of light by a substance (fluorophore...energy due to non-radiative energy losses. This difference in emission results in a slightly longer wavelength...phenomenon is known as the Stokes Shift, or the difference in wavelength between peak excitation spectrum...vary in their chemical structure, leading to differences in their excitation and emission wavelengths,...target specific organelles like mitochondria, and different dyes can result in varying cytoplasmic background...

  22. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ...Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers...bioluminescence resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite...problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently...Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent technology...need for an excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in ...technique’s versatility and adaptability to many different contexts. Improving Nano-lanterns In 2015, Takai...eNL constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition...

  23. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which... acceptor fluorophore. The difference for BRET is that the energy transfer is between a luminescent donor...number of different experimental approaches. They first assessed the activation of different members of...response to different agonists. These studies showed the promiscuity of GPCRs for different G protein transducers...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002...a vast number of signaling cascades, inducing different cellular responses. One of the biggest advantages...measure the physical association of combinations of different Gα, Gβ, and Gγ subunits. When inactive, these ...

  24. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...control many cellular processes such as differentiation, proliferation, and apoptosis. In humans, mutations...Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly luciferase. The bait and prey...variable availability of enhancer sequences in different chromatin states. Alexander Stark and colleagues... were deep sequenced to identify the gRNAs that confer protection to WNV. From this screen the authors...process of cloning, combining, and testing these different parts can be quite arduous. The cloning process...consists of a variety of plasmids containing many different promoters, RBS’, coding sequences (CDS), and terminators...To facilitate innovative cancer research, the Reference Reagents Group of the NCI RAS Initiative, led ...

  25. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ... variables.    Serotypes Serotype refers to the different protein compositions of AAV capsids, which... a virus enters the cell. Different serotypes utilize a number of different receptors, co-receptors, and...burdensome in the brain where many different cell types, or even different phenotypes of the same cell type...working in several different systems or want to transduce a large number of different cells. Promoters ...doi.org/10.3389/fnmol.2014.00076 References and Resources References Aschauer, D. F., Kreuz, S., & Rumpel...validation and optimization. Although there are different ways to ultimately achieve success, we find it...validation first.  AAV-mediated gene expression can differ extensively depending on the experimental system...

  26. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ... However, the two different approaches allow for optimized strategies for differing amounts of target ...urea-containing lysis buffer. Alternatively, the protein can also be eluted using a low pH buffer such as 0.1M ...for other antibody topics.     References and resources References:  Björck L, Kronvall G (1984) Purification... appropriate for most situations.  A slightly different method is desirable if the antibody is expected...Immobilization is the key to immunoprecipitation and simply refers to the process of anchoring an antibody, often ...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ...than those of agarose beads, despite the size difference. Magnetic beads are particularly advantageous...

  27. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...target primaries produced in different species and to label targets with different colors. Multiplex immunofluorescence...Speaking of different colors, having a palette of secondary antibodies in a few different species and ...dimensional epitopes. Different sample processing steps can alter epitopes in different ways, so antibodies... a successful IF experiment.    References and resources References Im K, Mareninov S, Diaz MFP, Yong...immunohistochemistry (IHC). All three of these terms refer to the use of antibodies to visualize a target within...essentially just more general terms that can also refer to assays using non-fluorescent visualization methods...indirect immunofluorescence There are a couple different approaches to be familiar with when talking about...

  28. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...blot Run your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize...before rinsing with wash buffer, as dry PVDF won’t evenly re-absorb aqueous buffer. Be sure to check the ...from having to run a whole new gel and membrane transfer. Let’s get started! Before you begin In the best...abundant. If the abundance of your targets isn’t too different, consider antibody affinity: stronger binding ...should not compare the strength of bands from different rounds of detection. Finally, stripping and reprobing.... Unlike PVDF, they can be returned directly to buffer without a re-wetting step. Important: do not dry...so you’ve dried your membrane and returned it to buffer, you’ve run and imaged your western blot with your...

  29. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ... IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation and had ...number, generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent... brain.  Figure 1:Schematic diagram of different FingR constructs and experimental results. Image... of the cell. By combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory... proteins system for Caenorhabditis elegans By Jennifer Tsang Fluorescent proteins (FPs) traditionally...The team validated this tool by targeting six different genes whose protein products have distinct localizations...Figure 2: Split-wrmScarlet labeling proteins with different subcellular locations. Image from Goudeau et al...

  30. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ...worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities depending... Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...make the virus. Viruses can be produced in many different cell types, spanning both prokaryotic and eukaryotic...However, some viruses can be produced in multiple different cell types, including plants, mammals, and insects...improve your current methods, understanding the differences can help you make an informed decision. Mammalian...AAV and lentivirus requires three plasmids — a transfer plasmid containing your gene of interest plus ...standard to do only one passage (transfection). This differs from typical insect cell preparations, which we...

  31. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...expression. Gene isoforms refer to mRNAs generated from the same gene but with different untranslated or coding...activated cell sorting (FACS) to isolate differentiated from undifferentiated cells at the end of the screening... to study neuronal differentiation, it could also be applied to the differentiation of other CNS cell ..., we’ll highlight these recent advancements and offer commentary on their application to neuroscience ...straightforward to use CRISPRa to selectively target these different transcriptional start sites in order to better... expression of Bdnf resulted in hundreds of differentially expressed genes between Bdnf I-overexpressing...which transcription factors promote neuronal differentiation (Liu et al., 2018). The pooled nature of this...

  32. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ... immune system make different isotypes of antibodies that recognize many different epitopes of the target... and protein G each have different affinities for IgG subtypes from different animals so it’s important...successful next experiment!     References and Resources References Ascoli CA, Aggeler B (2018) Overlooked... cover polyclonal antibodies, one of the many different types of antibodies available (others you’ll encounter...polyclonal antibodies from the serum There are a few different ways the antibodies can be isolated from the serum...by an animal are specific for the immunogen. A different type of purification can be performed to isolate...experiments where antibodies raised in several different species are present. One common example is using...

  33. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...Castle et al., 2014). Differences between AAV serotypes may be also linked to differences in viral uptake by...these different AAV vectors into a brain region, the AAV particles will infect neurons at different ratios...architecture, in particular the connections between different brain regions. For example, communication between...2015). Directional flow of information between different parts of the brain is mediated via individual ... for degradation. These two pathways also use different cytoskeletal machineries to facilitate transport...while anterograde transport relies on kinesin. Different rates of speed for various forms of retrograde... or have been genetically engineered to have differences in their capsid proteins. For a brief overview...

  34. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...Your AAVancement could be next!   References and Resources References Braun, C. J., Adames, A. C., Saur...allows for a wide variety of screens, including interference, activation, and knockout. Most CRISPR screens...CRISPR screens can be carried out using a few different methods — in vivo, in vitro, and ex vivo. The .... The go-to in vivo option AAV has become the preferred delivery method for in vivo CRISPR screens. AAV...titers equal to those achieved with lentivirus but offer an improved safety profile, customization, and enhanced...mouse tissues. Safety AAV and lentivirus have different safety profiles that often affect which is chosen... cause bad reactions in mice. Customization AAV offers a level of customization that is unmatched by other...

  35. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    Published
    May 23, 2023, 1:15 p.m.
    ...antibodies are all of different species/isotypes and that each secondary has a different color fluorophore...a comment below. Happy imaging! References and Resources References Andrews, N. P., Boeckman, J. X., ...be familiar with immunofluorescence (IF, often referred to as immunocytochemistry (ICC) when the sample...mixture of secondary antibodies, and image all the different colors in one session – fast and easy! Just make...Trimmer Lab, using recombinant antibodies with different isotypes. (A) Neocortex labeled with anti-pan-...make sure you are using primary antibodies from different species or isotypes, so that the secondary antibodies...to align the images of each labeled target as different color channels and analyze the results. Depending...

  36. Donations from Addgene to Yield Answers for Rare Disease Researchers

    Type
    Blog Post
    Published
    Sept. 15, 2015, 2:30 p.m.
    ...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies...will help them to understand how different mutations lead to differences in the symptoms experienced by ...competition for researchers working on rare diseases, offering them the chance to win some of the latest tools...with the condition to reveal some “outstanding differences.” They now have some theories on the underlying...had the opportunity to meet a few patients that suffered this disease,” Aguilar said. “Once you have that...Institute and Addgene will make a substantial difference in pushing the work forward. Xeroderma Pigmentosum...lot of those plasmids so that we can try many different things in parallel.” Alternating Hemiplegia of...

  37. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...demonstrated that different deaminase and UNG enzymes had different sequence preferences, which they defined...outcomes of their various CGBEs at different target sequences and in different cell types, available at www....non-target strand.” To avoid confusion, we usually use different terminology when discussing base editors, like...up to 68% at the most favorable target sites. Differential cytosine editing between bacteria and mammalian...sites. But the same strategy in E. coli produced different results. GBEs seemed to follow the same initial...editors Now that several labs had shown that preferential C → G editing was possible, the race was on ...creating BE variants with other Cas9s that recognize different PAMs. Their computational tool for predicting ...

  38. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ... with a T-DNA (transfer-DNA) region. During infection this T-DNA region is transferred into the plant....track, we switched to luciferases to more easily quantify the effect of the different treatments on transformation...method. Transfer your plant into the dark 16 hours before the transformation. We routinely transfer our plants...the T-DNA region with the gene of interest is transferred into the cells. But for C. roseus, Agrobacterium... lot of time. Don't re-invent the wheel. Test different leaf ages Usually younger leaves work better. ...GUS::tMAS to confirm transformation success. Luciferases are great quantitative reporters and use a substrate...substrate to produce bioluminescence. Luciferases usually have shorter mRNA and protein half-lives then...

  39. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...applicable to the rat [6]. Technical differences include the ferrule implants and patch cables. One huge...intracranial self-stimulation (ICSS) and place preference. The lab I work in (the Kreek lab) focuses on...behaviors to test reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting ...nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement but ...halves that are distinguished visually through different patterns and the animal only receives stimulation...). If a circuit is reinforcing, the animal will prefer to spend most of its time in the chamber in which...Comparison of c-fos amongst animals that receive different types of stimulation may help to refine and optimize...

  40. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ... transposition.  Target site: Different transposons insert at different DNA sequences or target sites....has several different applications (mutagenesis screens, transgenic animals, gene transfer). The discovery...specific locations in the genome.   References and Resources References Feschotte C, Pritham EJ (2007) DNA...commonly used in the lab While there are many different types of transposons, DNA transposons are most... vectors.   Let’s take a closer look at three different transposon systems adapted for use as research...Sandoval-Villegas et al., 2021). Sleeping Beauty’s preferred target site for integration is TA dinucleotides...piggyBac, Tol2 has a weak consensus sequence for its preferred target integration site: TNA(C/G)TTATAA(G/C)TNA...
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