We narrowed to 1,052 results for: FER

...yours does and then use that time! Different platforms have different quirks, so remember to practice on...that requires its students to do rotations in different labs before selecting an advisor, it is always...! Prepping for the interview Many universities offer free practice sessions for students doing virtual...points and keeping them at-hand, you can quickly reference your questions about the PI’s research or the ...the name of the award you won for your conference presentation. It can help soothe your nerves and make ...like sickness or scheduling issues can and do interfere with those in-person plans. Virtual options are...good fit for you. The application process looks different than it did a few years ago, but I hope the tips...

...extraordinary molecular detail.   References 1.Patterson, George H., and Jennifer Lippincott-Schwartz. "A photoactivatable...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...most scientists working on in vivo imaging models perfer the PA-RFPs over PA-GFP. Another class of irreversible...dimeric or tandem dimer versions of EosFP are preferred over the monomeric one since the formation of ...organelle trafficking. Monomeric PA-FPs are usually preferred for studying single protein characteristics, as... as the monomeric state is less likely to interfere with endogenous protein structure and function. By...non-invasive labelling, but in contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity...

...helping everybody, helps everybody! References and resources References Dayton, R. D., Grames, M. S., & ...scientists under-informed with respect to how different serotypes, promoters, and injection methods are...published.    Fig. 1: Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2...biological reagents; many factors, like storage buffers and conditions (3,5,8), injection titers (1,2),...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...& Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector dose–response and...sharing practices and data availability upon request differ across scientific disciplines. Scientific Data,...

...most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses...lentiviral vectors. Lentiviruses are sometimes referred to as “complex retroviruses ” (Cullen, 1992) because...generalize the idea of “transduction” to mean any transfer of material from the virus to the host. In fact...pseudotransduction” — to describe the direct protein transfer of RT, which would actually occur in step 2 during...dimerized linear ssRNA strands (which can have a different genomic sequence from one another) — by various... with lentiviral replication.  Resources and References More resources on the Addgene blog Viral Vectors...popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of ...

...to the plasmid search results: More than 200 different GCaMP containing plasmids are available for distribution...biosensor using AAV, you might decide you’d really prefer to order ready-to-use virus rather than producing...starting a new search or clicking through to a different part of the website, you can now use the pulldown...pulldown menu on the search page to quickly jump to different types of items: Did you notice the “Filter By.... The following screenshots show the range of different terms we can use to find materials from this publication...dropdown menus below the search bar. As you try different terms you may find the available categories and...you get this far and realize you need to apply different filters you do not need to start a new search....

... number of different strains of E. coli that can be used for transformation. The preferred E. coli strains...freezers that the samples can be stored in? Different shelves? Different racks on each shelf? Give each container...long-term storage in our facilities, every sample is transferred to a 2D barcoded tube (see next section) and ...about location, location, location There are many different types of samples at Addgene - DNA sent from depositors... concentration or need to be transformed into different cells than were originally requested. Most of ...analysis from our protocols and molecular biology reference pages! Resources Avaliable on the Addgene Blog...

... protein as pure as possible? Vectors that offer different combinations of tags are readily available,...peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant proteins...Purification Binding and elution steps use very moderate buffer conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK...co-IP, IF, flow -cytometry; can occassionally interfere with protein folding HBH HHHHHHAGKA GEGEIPAPLA...small size, rarely affects function S-tag KETAAAKFERQHMDS  1.8 Solubility and Affinity Abundance of charged...common. Tandem affinity purification (TAP) once referred specifically to a combo tag comprised of a calmodulin...information and some protocols can be found in the references provided. Additional resources: Young CL, Britton...

...delivered on a different plasmid and allows simultaneous multiplex editing of six different genes. This ... Jan 28, 2016 and last updated Sep 10, 2020 by Jennifer Tsang. CRISPR makes it easy to target multiple...flanking the promoter-gRNA construct, but with different sequences adjacent to the sites. When digested...assembly principles, but they’re optimized for different purposes.   Gersbach Lab multiplexing plasmids...kanamycin-resistant plasmids, each containing a different gRNA target sequence downstream of the 7SK, human...users to express up to 7 gRNAs. The kit contains different destination vectors depending on the total number...transcript. Rather than being transcribed from different promoters, the gRNAs are transcribed together ...

...between Cas9 enzymes from different species, explaining why they recognize different guide RNAs. Deep in the...Other CRISPR enzymes like Cas12 have different structures and different uses. In all cases, structural biology...techniques, like electron microscopy and FRET, can offer more information on these regions.   Cas9’s transition...from the same angle to highlight conformational differences. HNH, red; RuvC, orange; CTD, yellow; REC lobe...research. New details are still being uncovered, and different techniques all bring their own advantages—and ...Allergy and Infectious Diseases. Resources and references More resources on addgene.org  Addgene's CRISPR...656. https://doi.org/10.1038/s41579-022-00739-4 References Bravo, J. P. K., Liu, M.-S., Hibshman, G. N.,...

...translation of an array of different reporters including mScarlet-I, luciferase, antibiotic resistance markers...fluorescent proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor AC, Rees HA, Packer...et al., 2018. APOBEC-Cas9n-UGI targets its preferred trinucleotide motifs (5’-TCA-to-TUA) found within...using FACS which could limit their use across different cell types and systems. To introduce flexibility... GO works by affecting protein translation of different reporter proteins. Protein translation requires...Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image...

...lead to wildly different ideas of how to approach a problem, but it is these very differences that make our...the software design. Each of us brings slightly different cognitive biases as a result of our diversity ...Addgene and ourselves. We are also able to consider different technical approaches that may come up during discussion...developer and I get to see an approach that is likely different from what I would have done. And finally the stakeholder...isn’t how we work. We all take turns working on different parts of the code. I think the best way to learn...shipping sprint and all the stories will touch different parts of the shipping code. The following sprint...every part of the software, we are able to get different perspectives on how things are implemented and...

...immunotherapy than ever before. Although different types of cancer have different challenges and obstacles to overcome...However, there are many different forms of immunotherapy that work in different ways. For example, a new...learned much about the causes of cancer and the different avenues that can be used to treat it. For those...overcome, mesothelioma sufferers can see great promise in up and coming treatments like immunotherapy....latency period, and rarity of mesothelioma make it different from many other types of cancer. Mesothelioma ...mesothelioma, such as in the case of Paul Zygielbaum. A different approach to immunotherapy is bevacizumab (Avastin...

...husbandry and ability to produce abundant, externally fertilized, transparent progeny (Figure 2). These features...regulate processes from development to metabolism to fertility. Several zebrafish-specific gene expression databases...generating mutants. Unexpectedly, phenotypes sometimes differ depending on whether a gene is deactivated via ...researchers to observe changes to these tissues under different experimental conditions. Strategies for generating...tracing Over a period of 24 hours, the externally fertilized zebrafish zygote transforms into an organism ...sequencing approaches (Harland, 2018). Several different methods have been developed, relying on established...vertebratedevelopment using optogenetic approaches.     References El-Brolosy, Mohamed A., et al. "Genetic compensation...

...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills...graduate student at a networking event recently who referred to her lab as her family three times during a ...formulate hypotheses and conduct experiments. You transfer liquids between tubes, look through microscopes...etc.--so everyone’s experience will be a little different. Some of these changes you may enjoy immediately...production, I didn’t really start to recognize my transferable (or “soft”) skills until I was out of the lab...lab to think about these--get started with our Transferable Skills Guide.     Over the last decade, my... blog Visit our careers topic page Read our transferable skills guide Read blog posts about career readiness...

...Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding rep and serotype-specific...taken offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching...preparation undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products... matching system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding...After production, we confirm the identity of the transfer plasmid in the viral preparation by performing...performing PCR against unique regions of the transfer plasmid.   Lentivirus resources Browse lentiviral preps... by Ina Ersing in August 2023. Resources and references Resources at the Addgene blog Find and share ...

...tools with a focus on plants.     References and Resources References Karmakar, S., Panda, D., Baig, ...TnpB protein, much like how PAM sites change for different Cas species. For instance, the TnpB from Deinococcus...Deinococcus geothermalis (Dge10TnpB) targets a different TAM (5′-TTAT-3′). The unique TAM requirements ... genomic sites that can be targeted with TnpB, offering access to regions previously unreachable with ...commonly used nucleases like Cas9. This is where TnpB offers a clear advantage. At approximately one-third the...for plant genome editing using TnpB. The main difference between them lies in the choice of promoter for...Engineering the Plant Genome Using CRISPR/Cas9 RNA Interference in Plant Biology: New Tools for an Old Favorite...

...Bioluminescence Resonance Energy Transfer CRET= Chemiluminescence Resonance Energy Transfer    Interesting Articles...FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique which allows the detection of molecules...distance-dependent physical process by which energy is transferred non-radiatively from an excited molecular fluorophore...FRET experiments. You may have to design several different constructs to find one that could be used for ...higher than the signal generated by the energy transfer. On the contrary, if you choose a pair that are...cross-talk but you also don’t have any resonance transfer. You have to find a balance between FRET efficiency...RB, Periasamy A. Fluorescence resonance energy transfer (FRET) microscopy imaging of live cell protein...

...made it possible to offer DayCon as a free event! Organizing a science conference for the public How ...first-time event called DayCon. DayCon is a one-day conference aimed at the general public that provides an ...graduate student at Harvard, who wanted to provide a conference-style experience that was accessible to the general...How does one organize a public scientific conference? As a member of the DayCon Executive Board, I was fortunate...strength in the biomedical sciences and decided to offer a varied format event to keep the audience engaged... make DayCon a yearly event and to expand the conference to accommodate a larger number of attendees. ...

...written by Jennifer Tsang in October 2019 and updated by Emily P. Bentley in December 2024. References and ...(Anzalone et al., 2019). Plus, it can combine different types of edits with one another. All of this is...editing window cannot edit within what Fyodor Urnov refers to as “PAM deserts” in the genome. More versatile... instances where traditional base editors are preferred. For instance, if target nucleotides are positioned...type, and goal of the experiment can all impose different constraints and affect how you select and design...5.1-fold (though up to 45-fold) higher across different genomic sites compared to PE1 (Anzalone et al....used with any of the PE2-5 approaches, sometimes referred to as PE2max, PE3max, etc. PE6: Specialized prime...

...is to offer plasmids created as part of research projects, another way is to systematically offer thousands...worked closely with the legal partners and Tech Transfer Offices at universities to make COVID-19 research-related...United States, and many universities and Technology Transfer Offices were, at least, temporarily closing, these...scientists, we’ve worked with many scientists, tech transfer offices, research foundations, and funding initiatives...them available to industry, please reach out at techtransfer@addgene.org and we will initiate a conversation...conversation with you and your technology transfer office to discuss the possibilities for expanded sharing. ...

...sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different RTKs. Although...response. They generated variants optimized for different applications (v82 has higher affinity for glutamate...Measuring local temperature variations and heat transfer at the subcellular level requires a probe with...By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato...sub-millisecond resolution and sub-degree temperature differences under a variety of imaging conditions (155 μs...fast measurement of temperature and local heat transfer. Image reproduced from Lu et al. 2022, under CC-BY-NC-ND...plasmids here!   Lu K, et al. Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature...

...Addgene, I was about to have a check-in with my boss Jennifer. I had finally found my footing with the company... for what we thought would be a couple months. Jennifer and I spent about an hour that morning scrambling... involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs and benches...know cameras, but we needed someone else. Design: Jennifer thankfully volunteered, and we decided to focus...action would be to use an iPad to shoot.  Build: Jennifer created a mobile camera area that included iPad...Zoom, everyone’s new favorite third-party video conferencing software. Interviewees could simply boot up ...while the camera is here, the operator is in a different room.   Design: The previous cycle made me...

...by Addgenie Rebecca Hutcheson.   References and resources References Banasik, M. B., & McCray, P. B., ...second-generation system as an example.  The lentiviral transfer plasmid encodes your insert of interest, flanked...then, numerous variants have been developed with different mechanisms to inactivate the integrase.  The IDLV...vectors is random, and the integration site will be different for each transduced cell, there’s no telling which... that of regular lentiviral vectors. The only difference is that you would use the integrase-deficient...that titering these IDLVs may require a slightly different protocol than regular lentiviral vectors. This...quantifying the integrated viral DNA along with a reference cellular gene. When titering IDLVs, there is no...

..., protected samples and was compatible with the buffer additive MgCl2 and the crosslinking agent glutaraldehyde...PEs) via engineered virus-like particles (eVLPs) offers an option to transduce cells directly with ribonucleoprotein...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...

...solubility, and thus different reagents are useful for biotinylation in different microenvironments. There...used today in molecular biology for an array of different techniques and protocols. In this post we will...and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic acids to produce...relatively small, water soluble molecule that does not interfere with the macromolecules it is added to. Biotin... signal and easier detection. This strategy is referred to as the Avidin-Biotin Complex (ABC) immunohistochemistry...variety of biotinylation reagents available that differ in the functional groups or residues they target...a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular...

...scientists have developed and fine tuned many different ways to clone DNA fragments which have provided...more and more common, and for good reason. They offer many advantages over the traditional restriction...with an enzyme master mix, which contains three different enzymes: an exonuclease, which chews back the... this doesn’t allow for mix and match of many different fragments. One way around this is to use a combination...kit you will be using. Some of these products may offer advantages that Gibson assembly does not such as...with a G!) but actually work by significantly different methods. Golden Gate cloning does result in the...from one vector to another. There are lots of different ways to clone these days. Gibson and the other...

...are communicating across disciplines or doing different tasks. But these challenges can be exaggerated...equal opportunity to contribute. Using video conferencing tools like Zoom or Google Hangouts is a great...great way to include researchers from different locations. Having virtual face-to-face meetings as opposed... collaboration includes scientists working in different time zones, it’s important that everyone can access...project at the International Cancer Immunotherapy Conference in Frankfurt, Germany When funding and circumstances...techniques are being performed the same way at different sites. I did a lab exchange as part of the collaboration...been able to do from my home lab. Scientific conferences and workshops are another great place to meet...

...system which has been designed to perform gene transfer in vertebrates. The system is comprised of 2 components...transposase has been discovered in the genomes of many different fish species in a variety of non-functional forms...the lab showed that efficiencies of stable gene transfer using SB100x were comparable to stable transduction... vectors were developed into 16 variants with different selection markers (blasticidin, neomycin, puromycin... each contain two SfiI restriction sites with different overhangs depending upon whether they are on the...only likely to grow in the coming decades.  References 1. Kowarz E, et al. Optimized Sleeping Beauty... Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-...

...cells. The replicons of plasmids are generally different from the those used to replicate the host's chromosomal...include ColE1, pMB1 (which comes in a few slightly different but well known derivatives), pSC101, R6K, and ...regulated. Generally, control of replication is referred to as "relaxed" or "stringent" depending on whether...maintains about 20 copies per cell, while pUC – which differs by only two mutations – will produce as many as...considerations. *Requires pir gene for replication (reference). **F1 is a phage-derived ori that allows for...particles. Plasmids with phage-derived ori's are referred to as phagemids.   Other factors that affect...clarified? If so please, post in the comments.  References Chatterjee A, Sun SB, Furman JL, Xiao H, Schultz...

...out if you want to learn more. References and resources References Balloux, F., & van Dorp, L. (2017...called an envelope. Viruses without an envelope are referred to as naked or non-enveloped, while viruses with...viruses, here’s a quick tip on terminology. “Virus” refers to a single type of virus, not the individual viral...infection — researchers were able to create far safer viral vectors that can be used in BSL-1 or BSL-2...experimentally controlled conditions, making them far safer to work with than virions. The four types of viral...vectors (which many researchers will informally refer to as viruses) are named after the viruses they’...

...of many complex cellular behaviors, such as differentiation or metabolism. Our attempts to manipulate these...cerevisiae with few techniques that are easily transferrable across host species or metabolic pathways. A...system for multidirectional control The CRISPR interference (CRISPRi) model consists of a catalytically .... cerevisiae by using three RNA programs to differentially alter the levels of five genes that encode ...sequentially rerouted the metabolic flux to generate differently pigmented derivatives of violacein. Using dCas9...that will still recognize guide RNA but with a different dCas9 binding module, alternative sets of transcriptional...pharmaceuticals. Follow him on Twitter @AdamChinFatt     References Qi, L. S. et al. Repurposing CRISPR as an RNA-guided...

...activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase increases levels...transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and epigenetics ...delivery systems. Transcriptional repression DNA Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited...repression. Created with BioRender.com. DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1...Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase from the prokaryote Mollicutes spiroplasma...Schwiesow contributed to updating this article. References Haldeman JM, Conway AE, Arlotto ME, et al (2018... Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers...

...by Hannah Dotson in Dec. 2022.  References and Resources References Gossen M, Bujard H. Tight control...CMV, EF1A, and SV40, are always active and thus referred to as constitutive promoters. Others are only ...In the presence of tetracycline (Tet), TetR preferentially binds Tet over the TetO elements, allowing ...hear some transcription factor and promoter pairs referred to as binary systems. Binary systems are genetic... One can therefore place GAL4 and GAL80 under different promoters and create sophisticated patterns of...wondering, “why is it necessary to have all 3 of the different binary systems above if they all behave very similarly...s00253-018-9485-4. Epub 2018 Nov 3. PMID: 30392122. Pfeiffer, Barret D., et al. “Refinement of Tools for Targeted...

...they do arise. Happy CRISPR’ing! References and Resources   References Rabinowitz R, Offen D. Single-Base...cutting to occur. The RNA sequence and PAM site confer specificity of the nuclease; however, many Cas ...effects should impact your experimental design differently. Yes, it is always better to reduce off-target...mutation, off-targets must be quantified slightly differently. For a comprehensive approach, RNA-seq would ...to control for off targets as well as clonal differences. If you are performing CRISPR library screens... as a transfection or transduction control, a different gRNA would have distinct off-target sites, eliminating... there really aren’t any effective off-target reference controls. Off-targets and your experiment Every...

...For more context, exposition, and detail, please refer to the paper. Why might one be interested in state...register (Fig. 1C below). If attB-attP pairs from different recombinases are overlapping or nested on a register...that enters distinct states when treated with different combinations and orders of two recombinases. ...leveraging qPCR data from multiple primer pairs, one can infer the predominant state of registers within a sample...strong, directional terminators and promoters with buffer sequences on their 5’ and 3’ ends (for insulation... register. The purpose of this is to prevent interference between the transcribed gene and its 5’ UTR.... in biocomputing and biological circuits.     References 1. N. Roquet et al., Synthetic recombinase-based...

...pooled libraries are single preparations of many different plasmids. Plasmids within a given library have...the same backbone, but they express or target different genes. Some libraries cover the majority of the... library from the Moffat lab has over 175,000 different gRNA-containing plasmids). Amplifying and Using...screen. If you need to amplify the library, please refer to the depositor’s protocol for the best results...conducting pooled library screens. Addgene does not offer next-generation sequencing services. For an NGS ...Genome-Scale, Knockout Screening Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens...

...the liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have...cuvettes. First you measure the absorbance of the buffer that the DNA is in. This is a “blank” and measures...the ladder. You then run samples of your DNA at different dilutions and quantify based on band intensities...elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org...Addgene.org Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for ... Jennifer Tsang...

...in 1983. The differences between gram positive and negative bacteria is due to differences in cell wall...incredibly helpful in visualizing differentiations between bacteria, leading to safer medicine and more successful...minoring in studio arts photography.  References and resources References Silhavy, TJ., Kahne, D. & Walker ...negative and positive bacteria are permeable to different classes of antibiotics. The classification system...

... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...promoters or direct gene knockout. In contrast, CRISPR offers a much more user-friendly way to modulate gene ...Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting either Gram-negative...bacterial species (Peters et al., 2019). The systems differ on how exactly the CRISPRi locus is introduced ...Clostridium difficile, could be targeted and preferentially depleted from gut. While still far from the...the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig PM, ...

...updated by Jennifer Tsang in August 2020 and by Emily P. Bentley in September 2025. References and Resources...for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared to non-homologous end...APOBEC3G displayed less predictable sequence preferences.  The Liu lab also used natural and engineered...generation of base editors. Compared to BE3, BE4 offers a 2.3 fold decrease in C→G and C→A products as ...yielded an editor with more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more...the development of prime editing, which uses a different mechanism than base editing, has allowed scientists...Resources References Chen, L., Zhu, B., Ru, G., Meng, H., Yan, Y., Hong, M., Zhang, D., Luan, C., Zhang,...

... various AAV serotypes can be used to achieve different patterns of expression, researchers often need...empirically for their own experiments. To this end, we offer control AAV in smaller aliquots of 20ul for serotype.... The Addgene viral service has now expanded to offer many of our control plasmids in new serotypes, as...optogenetics lie in the spatiotemporal control offered by the system that allows for specific manipulation...H134R) bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression...Gordon Fishell   Chemogenetics Chemogenetics offers noninvasive methods for manipulating cell activity...dependent version of the DREADD hM3D is now also offered as AAV1 (along with AAV PHP.eB, AAV Retrograde,...

... scientific conference, whether you are attending in person or monitoring that conference via social media...Fast Company, opportunities to speak at conferences, and offers for postdoc positions. Check out the podcast...write a second. While working on this post, I was offered and accepted a position to work remotely as an ...American Society for Cell Biology (ASCB) 2018 Conference. I don’t exactly know how I was selected to participate...s worth a read since many of the pointers are different from the other links shared in the post. And the...

...new features to the tool. Resources and References References Adler, B., Trinidad, M., Bellieny-Rabelo...solve the problem: CasPEDIA, a new resource from Jennifer Doudna’s lab at the Innovative Genomics Institute...CasID, to help facilitate direct comparison of different enzymes. CasPEDIA is meant to be a constantly ...plasmids containing that enzyme, and embedded in the References lists on every page you can find direct links...design, etc.). With this feature, you can compare different Cas enzymes to make sure you are getting the desired...the winner!).   Fig. 3 – Comparison of different Cas enzymes using CasPEDIA. Adapted from Adler...

... 2019), and different combinations of ITR and capsid serotypes vary in their gene transfer efficiency....DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs,...and rAAV vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et al...expression can be altered by using ITR sequences from different AAV serotypes that vary in their RBE and T-shaped...restriction digest methods.    Resources and references More resources on the Addgene blog Viral Vectors...Addgene.org Addgene's AVV guide AAV Viral Preps References  Chen, Y., Hu, S., Lee, W., Walsh, N., Iozza...

...and it’s already looking like it’s making a big difference. We can now look at many more transcription factors...Mendenhall: The system is really quite simple but it offers improvements in efficiency and throughput. In adapting... this for tagging any protein in all kinds of different assays, not just transcription factors. The second...quite small—20 amino acids—so we don’t think it interferes with the function of the protein. Myers: We’ve...Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks... surprising to us that there aren’t a hundred different versions of this out there. Addgene: Why was it... They could also switch from FLAG to GFP or a different epitope. Addgene: Do you have any additional tips...

...delivery of split base editors Article contributed by Jennifer Tsang AAVs are a great gene therapy delivery mechanism...reproducibility as scientists can assemble parts from different labs and experimental systems in their own research...mutants, determined protein expression levels from different promoters, and inhibited gene expression using...contributed by Erin Sanders CreLite, designed by the Eisenhoffer lab, is a new tool that increases temporal and...bringing the two halves of Cre together. Image from Eisenhoffer et al., 2019. This novel tool is practical...organ culture, as well as cell lineage studies. Eisenhoffer and his colleagues evaluated CreLite for cell...eye, muscle and epithelial cells, as well as in different stages of development, ensuring the breadth and...

...al. designed plasmids containing Cas9 and two different gRNAs, and electroporated them with an ssODN repair...experiments in S phase zygotes at 18 hr post fertilization. They observed higher targeting efficiency than...41/42 embryos were non-mosaic at 3 days post fertilization. Since Ma et al. designed a repair template ...about the creation of “designer babies.” The differences in editing between iPS cells and embryos make...off-target effects, but this advantage likely won’t transfer to all genes we’d like to target. In addition,...embryos to only 60%? Unfortunately, given the differences between CRISPR repair in embryos and iPS models...available with current techniques like PGD.  Reference Ma, Hong, et al. "Correction of a pathogenic gene...

...Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different plasmid, which may lead... entry vectors (pUC57-L1L4 and pUC57-L3L2) that offer the advantage of using restriction digest cloning...plasmids to study PPI using Förster Resonance Energy Transfer (FRET, Heckler et al., 2015) or ratiometric Bimolecular...consists of 4 plasmid toolkits generated with 4 different, binary 2in1 vectors with all possible combinations...change in protein conformation in response to light, offers a way to spatially and temporally control cellular... or dark- state of the photoswitchable proteins offers a powerful approach to control protein-protein ...

... to what’s happening at different points within the wood. There are different types of cellulose fibers...number of DNA binding proteins with Cas9 guided by different promoters. The challenge in building genetic circuits...getting enough regulatory proteins that don’t interfere with each other. CRISPR is so orthogonal and programmable...then you’d need the ability to turn on genes at different times. It requires more complex synthetic networks...circuits, they are extremely orthogonal. It’s a different perspective and you can layer them for more complicated...for multi-input CRISPR/Cas9 genetic circuits References: Nielsen AA, Voigt CA. Multi-input CRISPR/Cas...

...numerous variants for the different types of opsins available and their differences and uses have been discussed... the different variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin...about this in detail later). But what are the different steps in an optogenetics experiment and what are...available, instructions for obtaining a material transfer agreement, and list of vendors (such as the vector... core at the University of North Carolina) that offer his constructs. Importantly, control viruses (not... or follow him on Twitter @derekpsimonphd.    References 1. Tye KM, Deisseroth K. Optogenetic investigation...