...Localization Determine where your protein of interest resides by using a well-characterized fluorescent fusion... Medical School Fluorescent Biosensor Database designed by Eric Greenwald and the Jin Zhang Lab at UCSD...

...the order of nucleotides in a string of DNA (7). Sanger sequencing uses modified nucleotides, called dideoxynucleotides...the widespread use of Sanger sequencing. Building upon this technology, radiolabelled nucleotides were...causes sickle cell anemia. The mutation happens to destroy a sequence recognized by the restriction enzyme...synthesis reaction along with all four regular nucleotides results in the synthesis of DNA strands of all...fluorescence activity as fluorescently labeled nucleotides are incorporated, a method called pyrosequencing...nanopore-based DNA sequencing which reads the nucleotides of a DNA strand as it slips through a tiny pore... the different electrochemical properties of nucleotides (8). Compared to older technologies nanopore-...

...Have you ever designed a CRISPR guide RNA and wondered why it is limited to only 20 bases, or why it’...understand the parameters of CRISPR experimental design. Let’s dive into some structural biology! Major... lobes, REC and NUC, and their subdomains. NUC includes the HNH and RuvC catalytic domains, as well as...as the CTD (also known as the PI), while REC includes most of the bridge helix. Created with BioRender.com...Fact! Even though the Cas9:RNA complex is well described in the closed conformation, recent electron microscopy... does not include stem loop 3, and guide RNA nucleotides 11-20 are not resolved due to structural disorder...2017). Within an interior cavity, Cas9 grips nucleotides 1-10 of the guide RNA, prearranged for base pairing...

...larger Antibody Plasmid Collection , which also includes other types of affinity reagents such as scFvs...Browse All Antibodies ID Recombinant Antibody Description Source Species Isotype Reactivity Recommended...

...Plasmids Available as Cloning Grade DNA ID Plasmid Description Industry PI FAQ What Quality Control analyses...cloning grade DNA into cells? Cloning grade DNA is designed for use in cloning applications. Direct transfection...

...Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in various serotypes suitable... Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV in various serotypes suitable...

...10.1038/nprot.2009.98. Epub 2009 Sep 17. PubMed . Description Use of the OPEN method requires OPEN pools (available...using the bacterial two-hybrid system originally described by Hochschild and colleagues ( Dove et al., Nature...

...from Wong, 2018   Primers Design of primers and probes Primer design, of course, could easily be its...useful primer design resources in the References and Resource section. Validation Once designed, your primer.... For example, chemistries that rely on hairpin design of probes/primers often offer higher allelic specificity...Taqman) Low; can be reduced with a double-quencher design Yes, one Yes Yes Agarose gel or uMelt analysis...Reference Genes  Choosing a set of reference genes deserves more attention than it usually receives. Different... multiple reference genes whenever possible (Vandesompele, 2002), and it's possible to multiplex your ...documented in the literature, a geNorm study (Vandesompele, 2002) will help you select the appropriate ...

...this article, Quilt CTO Aneesh Karve, shows how to design experiments that work anywhere in the genome. Aneesh's...enhancers! This leads us to a three-step approach for designing an experiment to identify the critical NANOG binding...Find Enhancers that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG...anyone can find enhancers with Quilt. Seek and destroy enhancers with CRISPR Armed with the genome math...expression for NANOG enhancers, we’re ready for step 2: design a CRISPR screen to disrupt these enhancers. The...attenuated retrovirus in the same family as HIV. By design, our lentivirii are genetically programmed to CRISPR...Lentivirus from Addgene If you’re interested in designing your own CRISPR screens for enhancers, check out...

...many MoClo kits are designed with specific model organisms in mind, others are designed for flexibility. ...Synthetic biology in bacteria MoClo is commonly used to design TUs and multi-TU constructs for synthetic biology..., biofuels, and natural products without the undesirable side products generated during chemical synthesis.... phaffii, then the MoClo YTK Kit specifically designed for S. cerevisiae is at your service (Lee et al...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within... are endless: genetic circuits with multiplexed guides, mini screening libraries, direct edit optimization.... The Fragmid kit described above works in both mammalian systems and Drosophila. Similarly, the Multi-Kingdom...

...their specificity or from the thymus to the lymph nodes. These methods gave researchers much more information...expressing cells. For example, by harvesting lymph nodes at different times after antigen challenge, scientists... cellular dynamics in lymphoid organs. In lymph nodes, for example, two-photon microscopy allowed scientists...extravasate from the bloodstream to invade lymph nodes and scan antigen presenting cells, but two-photon...interactions in diverse tissues, including lymph nodes, brain, liver, gut and skin. Two-photon intravital... 2016)? How do dendritic cells migrate to lymph nodes to activate T cells (Celli et al 2008, Kitano et...dynamics within the germinal center of murine lymph nodes. However, one must pay attention to time scale when...

...affects all kingdoms of life. Of course, that includes you. Besides the risk to the researcher, Zeocin has been...most yeast vectors and many mammalian vectors are designed to shuttle between species. Figure 1: Shuttle...host’s replication machinery. A shuttle vector designed for unrelated bacteria should include oris for...target species. Fun fact! Plant vectors are usually designed to shuttle between E. coli and Agrobacterium tumefaciens...protects against any of them. But different aminoglycosides work on different host species: kanamycin targets...media that supplies the key nutrient. The vector provides the missing synthesis machinery, and the cells...image for individual analysis. A shuttle vector designed for this approach in a mammalian cell experiment...

...reproducibility than others. Unfortunately, most ELNs are designed as closed ecosystems and act as data silos because...RSpace ELN supports reproducibility RSpace was designed in response to requirements that emerged from ...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates... in institutional and lab servers specifically designed to hold large amounts of data. Although it may... use of other tools they use in research. This includes chat apps like Slack, Microsoft Teams, and Google...individual documents or more complex datasets into the desired repository. They also support entry of required...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...

...Patrick Miller-Rhodes from the University of Rochester Medical Center. Patrick Miller-Rhodes is currently...This post was contributed by Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at ...this technological gap, a trio of recent papers describe the development of CRISPR activation (CRISPRa)... large size of Cas9 activators, which largely precludes delivery by adeno-associated vectors (AAV). This...packaging capacity than AAV. Such an approach was described recently by the Day Lab, who developed neuron-... selected the immediate-early gene Bdnf, which encodes brain-derived neurotrophic factor (BDNF). Bdnf ...High-throughput screening strategies Although the tools described above make CRISPRa more accessible to neuroscientists...

...property by including ccdB on its destination plasmids. In the destination plasmid, the ccdB gene is flanked... rest do not grow. While there are many ways to design your positive selection strategy, here are a few...straightforward in cases where the inserted DNA includes an antibiotic resistance marker. After transformation...grow. However, often times, the plasmid backbone includes an antibiotic resistance cassette and you aren...backbone contains your gene of interest. Auxotrophy Besides antibiotic selection, you can select for a successful...prototrophy come into play during cloning if you’ve designed your experiment using an auxotrophic strain of...sites. To introduce a gene of interest into the destination plasmid, recombination sites must first be added...

...Aptamer Fluorophores describes the enrichment process used to evaluate oligonucleotides for binding affinity...of Spinach. In doing so, they identified the nucleotides that contribute to the instability of Spinach...similar levels of fluorescence when bound to DFHBI despite being approximately half the length (Warner et ...also smaller than any form of Spinach - only 49 nucleotides in length - and is better able to maintain its... of colors, so do aptamers! Corn was recently described in 2017 (Song et al., 2017). Corn naturally dimerizes... Red and orange fluorescent aptamers Corn was described in the same study as Red Broccoli and Orange Broccoli...Broccoli, and share similar properties to Broccoli despite binding to a different small molecule. Notably,...

...for the desired modification. Finally, the positive clones are sequenced to confirm the desired modification...Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the desired mutation...introduced to plasmids using primers (with the desired mutation) in a PCR protocol that amplifies the ...mutated clones. Forward and reverse primers are designed to be complementary, but each primer may extend...Polymerase To ensure that you don’t introduce undesired mutations through the PCR process, you need a ...The resistance marker from the parental plasmid provides a mean for selecting for transformants which have...mutagenized plasmids Once constructs containing the desired modification have been identified (and confirmed...

... Addgene plasmids by gene here), designing primers with your desired mutation, performing PCR to generate...multiple fragments of DNA to create a synthetically designed DNA sequence. There are multiple methods of DNA...purification steps and end-repair steps with well-designed fragment overlap sections, while still creating... guide you through both experimental and primer design based on fragment sequences and the polymerase ...enzymes are adjusted. Appropriate fragment and primer design results in covalently joined, scarless DNA – perfect...by assembling multiple fragments containing the desired mutations. This same process can be used simultaneously...oligo that contains a region of homology and any desired edits to the target region. Assembled plasmids ...

...express a gRNA designed to direct Cas9 to their target sequence of choice (see tips for designing a gRNA) and... of the major CRISPR lingo and will be able to describe the functions of the various CRISPR/Cas9 components...responsible for processing foreign DNA as well as the RNA guides required for endonuclease function, when used for...Bind to a guide RNA The guide RNA, which will be described in more detail below, enables Cas9 to cut a specific...DNA damage is a DSB within the target DNA ~3-4 nucleotides upstream of the PAM sequence. Cas9 Species/Variants...with the appropriate PAM sequence) to modify the desired genomic locus.   Some historical notes CRISPR ... combined with its adjacent spacer, ultimately encodes a single crRNA. The direct repeat regions contain...

...diversification of all nucleotides at a user-defined loci in bacteria. Its modular nature provides versatility in...provide a means to continuously diversify all nucleotides within a user-defined genomic region. EvolvR,...method developed by the Dueber Lab at Berkeley, provides a new approach for generating novel genetic variants...users to achieve targeted diversification of all nucleotides within a defined region of a genome. Require...turbidostat. Only mutate particular types of nucleotides. Both the Bassik Lab and the Chang Lab have created...3’ end, while its native endonuclease activity degrades the displaced strand.   Figure 1: EvolvR...single starting cell. Additionally, multiplexing of guides allows EvolvR to target more than one genomic site...

... error rate for long sequence knock-ins Visual barcodes for live cell clonal multiplexing Improving ...nanobody-GFP interaction). The modularity of the system provides researchers better flexibility and makes the approach...other experiments. And, while the BLITZ system was designed and optimized with zebrafish in mind, it can easily...https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal multiplexing by: Rachel Leeson...Recently, Ravid Straussman’s lab has developed visual barcodes for clonal multiplexing of cells in live imaging...peptide. By combining three different localization peptides with four different fluorescent proteins, and ...freeware.    Fig. 4: Images of 12 visual barcodes in use in A375 cell line. Image from Kaufman, ...

... is also documented. Experimental design When you are in the design phase of your experimental planning...CasPEDIA is an online, searchable resource that provides summary information on Class 2 Cas enzymes. The...The information is organized in wiki format and includes biochemical properties, previously established...relevant resources The homepage displays a graphic describing CasIDs and relevant properties of Cas enzymes...What other RNA components are required? CasPEDIA provides information on crRNA and tracrRNA requirements... also show you how multiplexed arrays would be designed, if that’s your end goal. CasPEDIA also features...search (trans activity, multiplex ability, gRNA design, etc.). With this feature, you can compare different...

...site by the pegRNA — a guide RNA that also encodes the desired edit and homology to the genomic DNA locus...both specifies the target site and contains the desired edit (or edits) engages the prime editor protein...and the reverse transcriptase domain copies the desired edit, directly polymerizing new DNA onto the nicked...edit within what Fyodor Urnov refers to as “PAM deserts” in the genome. More versatile and precise than...editors are preferred. For instance, if target nucleotides are positioned within the canonical base editing...still almost always results in higher ratios of desired edits to indel byproducts than Cas9-initiated HDR...different constraints and affect how you select and design your prime editing tools. Here, we’ll briefly cover...

...Voigt Clifford Wang Ron Weiss Synthetic biology describes the rational engineering of biological systems...Phosphoprotein Kit S. cerevisiae Advanced Gateway Destination Vectors TAL Effectors Zinc Finger Consortium ...

...A recent publication [1] from the Sánchez lab describes the plasmids and protocols required for both WISH...companion resource called “cutting class” which provides more detailed materials and protocol resources...synthetically made RNA (riboprobes) with modified nucleotides (for example, Digoxigenin-labelled Uracil) that... molecular cloning to insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow... which are available through the deposit) are described in the Sánchez lab publication [1] so that the...expressed mRNAs.These native mRNAs are subsequently destroyed by Argonaute-catalyzed nuclease activity or their...have to use a commercial E. coli strain that is designated as DE3. For Phagocata morgani and Phagocata gracillis...

...cloning, or MoClo, is a systematic cloning strategy designed by synthetic biologists to perform large scale...specifically refers to the organizational schema described above, and not any particular cloning technique...example of an individual part, this category also includes promoters, UTRs, terminators, etc. Once selected...behave in a cell. For unpredictable cases, MoClo provides two answers: first, sets of parts with characterized...protein products can interact with the DNA that encodes the protein itself or other genes within the circuit...cellular engineering goal, several of which are described below. Multigene circuits assembled through MoClo...for more complex CRISPR experiments (e.g., many guides/parts delivered) in certain model organisms. In...

...envelope and packaging plasmids. ID Plasmid Type Description PI 8449 pUMVC Packaging Contains MoMLV gag and...plasmids that have inserts. ID Plasmid 5'LTR Description PI 1780 pBABE-neo largeTcDNA MoMLV Expression...

...for and how can you do it well? Below, I briefly describe some tips for a successful postdoc fellowship....typically writes the grants that fund your research, guides the research directions of the lab, gives you constructive...Thesis committee members provide expertise in designing and interpreting experiments, as well as career... there a tractable question or are you simply describing a phenomenon? Exploration can be important, but...presentations benefit from practice. If you simply recycle slides from the last time you gave a presentation to a...story I can tell with my data?  Can I make better slides that more effectively tell a story? Generally,...or functional models or more helpful background slides. If you don't have much new data, speculate on ...

...extremely useful if a desired protein product is toxic to cells at high concentrations. Despite their advantages...and induced by lactose, provides the highest level of protein expression. Despite their ease of use, it...for easier access and purification of a desired protein. Despite being a rather nascent technology, plant...for proper protein function. Most DNA constructs designed for mammalian expression utilize viral promoters...protein in bacterial cells: the pET, pRSET, Gateway pDEST, and pBAD vectors for example. Protein expression... include ribosomes, RNA polymerase, tRNAs, ribonucleotides and amino acids. Cell-free expression systems...

...Plasmid purpose: This is a brief description of what the plasmid was designed and used for in the depositing...plasmid, how will it be sent to me? Description: This describes how the plasmid or other product (kit... Gene/insert: This describes what has been inserted into the backbone, and provides information about ...successful cloning. Cloning information: This section provides useful information about how the vector was assembled... sequencing. Growth in Bacteria: This section describes how to grow the bacterial strain in which the ...relatively new feature of our plasmid pages, and provides a list of publications which have used the plasmid...

...label of their choice according to the methods described by George et al. (2004). The ACP and SFP Synthases...respectively) may be expressed and purified as described previously in Stachelhaus et al. (1998) References...

...reporters are challenging to design and difficult to multiplex. The Covert Lab provides an alternative tool with...fluorophores can be multiplexed together. Challenging  to Design: FRET reporter activity depends on both the distance...advantages. First, they are easy to multiplex. Their design allows flexible swapping of fluorescent proteins...overlap are used together. Second, KTRs are easy to design. The key determinants of KTR function are the positions...See Box 1 in Kudo et al. for more details on the design of the bNLS and NES sequences. Disadvantages of...translocation rates are another variable to consider when designing KTR experiments. Third, KTRs are not well suited...JNK, and ERK, or a backbone to use for custom KTR design, plasmids are available from Addgene! References...

... Zimmer provides several. In Part Three of the book, called simply “Old Science,” he describes the now...teaching and communicating science for over two decades, attempts to answer it in five paragraphs, with...recent resurgence of “Do-It-Yourself Science” described in Chapter 3, and of course the rapidly changing... science.”  The three chapters in this section describe the challenges of trusting science in a “fake ...changed significantly over the last decade. He describes the detection of gravitational waves--a story ...collaboration from scientists around the world. The paper describing the waves’ detection had over 1,000 authors. ...This last section of the book, Future Science, describes the author’s thoughts on science’s biggest challenges...

...These libraries utilize optimized sgRNAs that were designed based on the rules identified in Doench et al....1,000 non-coding control sgRNAs. This library was designed to improve on-target activity while reducing off-target...with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello library... that if an essential gene plays a role in the desired screening phenotype, it may be missed because it's... the sgRNA metrics, confirming again that sgRNA design plays a significant role in library efficiency....CRISPRa, sgRNAs were targeted between 150-75 nucleotides upstream of the TSS. Similar to the Brunello ...Activation Mediator) uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with...

...Researchers have used oligonucleotides in fluorescent visualization techniques for decades, most notably for... exponential enrichment (SELEX), some 1010 oligonucleotides could be screened for their affinity to a ...aptamers as well, and over the course of the next two decades, these nucleotide-based ligand binders would prove...their broad range of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells (even...counterparts, antibodies and enzymes, such as: Design and selection for targets can be done largely in...such techniques can be problematic, however. Well designed aptamers can help reduce such background, thereby...relatively bulky. One of the advantages of oligonucleotides over proteins is their modularity, which is...

... about! You could describe your organizational system, your rationale for designing it the way you did...experience and skills would help you do the job as described. Let’s imagine you’re applying to a job that ... your cover letter, you can expand on that by describing how you developed and used a ddPCR assay to answer...question. Now, the hiring manager knows that you can design experiments with ddPCR, interpret the results, ...start by looking at the requirements in the job description. For each requirement, ask yourself if you have...to their work. Since the beginning is often the hardest part, here are some examples to get you started...company/organization]. I am a [adjective that describes you] scientist with over [number] years of experience...

...vectors and design your sgRNA targeting sequence using one of the many freely available sgRNA design programs...many cases several exons’ worth of DNA. Any ∼20 nucleotides of DNA can serve as the target for Cas9’s single...Cas9 localizes the deaminase to where the edit is desired, and it performs the programmed edit within the...flexibility. A consideration with this tool is that the desired edit will be performed on all relevant bases within...system utilizes an RNA template containing the desired edit adjoined to the sgRNA, a reverse transcriptase...not require an expertly positioned PAM without undesired bases nearby that may undergo unwanted editing....   Ok, I have a Cas9! What do i do next? Designing or selecting a sgRNA Addgene may carry a validated...

...contamination and match their original description. The oldest, largest, and most diverse of these repositories...culture in the present day Cell culture is a widespread tool used in the fields of oncology, virology...problems surrounding cell culture over the last few decades some pharmacologists have tended to avoid cell ...International Cell Line Authentication Committee includes a database listing all cross contaminated or misidentified...comparable studies to be performed on a cell line even decades after it is initially isolated. Regrettably this...Master and Working stocks to work from. The future Attitudes are changing but yet more is needed to convince...can be prevented and we can progress faster to desired clinical endpoints.   Nick Amiss is an ATCC Outreach...

...create and express plasmid-based antibodies. Description Article PI One-step assembly of antibody heavy...for "anti-mouse" or "anti-rabbit") ID Plasmid Description Gene/Insert PI Return to top Do you have suggestions...

...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific... tool for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring ...iGECInano plasmids here!   Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared...intermediate affinities from 40 to 140 mM. These new designs greatly broaden the options for potassium biosensors...further engineering. Figure 5: Structure-guided design and genome mining both led to new sensors with ...

...Good as a WFH desk, and as a video control center.   Cycle 2: Zooming Forward Design: We produced ... So, much like SynBio, it was time to start our Design-Build-Test-Learn cycles. If you’re not familiar...again. Here’s that process!   Figure 1: The Design-Build-Test-Learn cycle adapted for video making...through. I know cameras, but we needed someone else. Design: Jennifer thankfully volunteered, and we decided...here, the operator is in a different room.   Design: The previous cycle made me realize we needed to...restrictions in place, most importantly, masks.  Besides the muffled voices beneath masks, humans rely on...reach for Addgene.  So, onto research! I needed to design a system that would allow me to set up a camera...

...suggests there are hundreds. This only includes general lab strains designed for subcloning or protein expression...was updated on Nov 14, 2017. You've worked hard designing your plasmid – you carefully selected the optimal...Dr. Theodor Escherich, the scientist who first described them in 1885. E. coli are mainly found in the ...commercial lab strains of E. coli used today are descended from two individual isolates, the K-12 strain ...mutations found in E.coli strains Gene(s) Description Functional Consequence dam DNA adenine methylase... fertility plasmid. A low copy-number plasmid, encodes proteins needed for bacterial conjugation. Genes...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular strains...

...please improve your plasmid maps!" After thoughtful design, vetting, and tweaking, we’re excited to announce...identifies plasmid features that differ by only a few nucleotides, such as the fluorescent proteins EGFP and mEGFP... software. In fact, SnapGene’s feature library includes nearly 3x as many features as could be detected...greater than our previous mapping software and now includes Type IIS restriction enzymes that are commonly...the gene, the name of its gene product, and a description of its function. You can also click one location...enzymes that cut the given sequence. The table includes enzyme names, the locations where the enzymes ...features detected in the plasmid sequence. The table includes feature names, locations, sizes, colors used to...

...have pre-designed plasmids for these systems! Ion sensors One common biosensor class is designed to monitor...all have specific sensors designed for them. For example, this class includes tension monitors associated...interest for many disciplines! Most of the sensors designed for calcium generate a fluorescent output, which... fluorescence in the presence of calcium. This includes FRET-based sensors such as Twitch. While calcium.... Physiological factors Unlike the previously described sensors, this class measures either factors as...to use them, you’ll need to use the vectors (or redesign the tools). It’s a good thing, though! Viral delivery...

...discussed methods. As described above, you design two primers, each containing the desired mutations, to amplify...make a point mutation, you design two oligonucleotide primers with the desired mutation, then use those ... figure above shows guidelines for primer design; a desired insertion should be present in both primers...vitro recombination. Across all modifications described in Trehan et al., REPLACR-mutagenesis’ median ...

...recombinant protein purification and cell-free circuit design in E. coli - Freemont Lab MoClo-YTK : Assembly ...resources, such as depositor protocols and gRNA design software. Addgene yeast CRISPR resources Bacterial...

...coveted plasmid with no substantial delays. Design, design, design Before diving into the experimental work...experimental work. Fragments generation After you’ve designed your cloning scheme, you’re now ready to generate... to use 1-2 ug of the vector for digestion. PCR Design primers that overlap at least 24 bp with the template...from a large genome, you can recheck your primer design or optimize your primers or PCR conditions. Purifying...second and more detailed look at your experimental design. Full lawn of cells. The antibiotic in your plate...complementary to standard sequencing primers. If not, design primers to sequence your insert. When you get your...

... model to help them construct “designer” channelrhodopsins with desired characteristics. The model was...Viviana Gradinaru’s lab used machine learning to design new channelrhodopsins that are more light sensitive...constrained and poorly understood. It’s hard to design new channelrhodopsins with high-throughput proteins...Using machine-learning to overcome channelrhodopsin design challenges To overcome these challenges, the Arnold...experiments. The sequences in this library were designed using structural information about parental channelrhodopsins...existing channelrhodopsins CoChR and/or ChR2(H134R). Designer channelrhodopsins paired with a systemic AAV allow...Find our chemogenetics and optogenetics science guides ...

...utilizing chemogenetics (Designer Receptors Exclusively Activated By Designer Drugs, DREADDs) and optogenetics...beg the question: is axonal transport desirable or not desirable? If diffuse gene expression is tolerated...examining the use of DREADDs (Designer Receptors Exclusively Activated By Designer Drugs) to determine the ...to their Biosafety Level-2 or 2+. Please Note: Despite the fact that AAVs themselves are not pathogenic... CNS tropism (4). If axonal transport is not desirable, transport can be reduced by injecting a smaller...miRNAs), short hairpin RNAs (shRNAs), antisense oligonucleotides (ASOs), zinc finger nucleases, and many others...

...Marraffini’s lab took advantage of this lethality to design the first synthetic bacterial CRISPR system in ...gRNA cassette for plasmid curing through "self-destruction" after induction with L-rhamnose and aTet. CRMAGE...citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while pCRISPomyces...editing efficiency, perhaps due to its condensed design. For both systems, custom spacers/gRNAs are easily...et al., 2013), the Qi lab employed a gRNA-based design more familiar to those using CRISPR in eukaryotes...plasmid tools that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab...