We narrowed to 872 results for: fer

...cloning into a modular cloning strategy. Sometimes referred to as MoClo, this strategy uses the Type IIS restriction...256 distinct flanking sequences, sequences that differ by only one base may result in unintended ligation...constructs, assemble standard plasmids parts in different combinations, or other new and exciting applications...constructs in a single, high-efficiency step. References: Sequential amplification of cloned DNA as tandem...

...CRISPR experiment. Happy cloning! References and Resources References McGee, A. V., Liu, Y. V., Griffith...chosen because of their broad applicability in different model systems. What’s the error rate? The Doench... pRDA_722 (CMV-driven; lentiCRISPRv2-based; preferred over 512) Mammalian Lentivirus pRDA_789...

...and epigenetics. So, I sought out a number of different opportunities to broaden my training beyond the...major part of our everyday work is to liaise with different organizations and individuals, to identify ways...collaborating, and to organize events that bring different voices together to talk about genetics. In addition... such as scientific, educational or industry conferences, as well as schools, community events, churches...

...more inputs than was previously thought. The differences in the inputs to each of the two populations ...populations could be used to explain the differences in the functions of these two neuronal populations. Overall...Overall, this study laid the foundation for how different anatomical regions of the brain receive inputs and...complex questions of how the brain operates. References 1. Callaway EM. Transneuronal Circuit Tracing...

... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover...changes in TagRFP-T fluorescence intensity at different time points, the authors tracked protein activity...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy... FLIM-FRET acceptors here. See Addgene's FRET reference page for many other common FRET vectors. Murakoshi...

...28032891. This is a different version of Split-BioID. BirA* is fragmented at a different site than in Schopp...of the BirA* fusion matters! Schopp et al saw different levels of fusion protein expression and biotinylation...research? Plasmids are available from Addgene! References Schopp, I.M., Ramirez, C.C., Debeljak, J., Kreibich...

...StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific AAV delivery...quickly saw wide interest, its dimeric nature can interfere with some applications. Several labs have been... viral vectors for functional intravenous gene transfer throughout the non-human primate brain. Nature...the Gradinaru lab, AAV1-X1, was developed by transferring the 7-mer peptide region from variable region...

...more precise editing in the lab. References and resources References Dong D, Guo M, Wang S, Zhu Y, Wang... May 23, 2017 and last updated Jul 23, 2020 by Jennifer Tsang. CRISPR-Cas technology is constantly evolving...loadingExample: AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas...

...compatible with multiple FP(11) fragments, even from a different base FP (for example, EBFP2(1–10) can be used ... useful. Good luck, and happy experimenting! References and Resources Additional resources on the Addgene....org Fluorescent Biosensors Collection Split Luciferase systems, including NanoLuc®, DULIP, LuTHy Other... Fluorescent Proteins Plasmids and Resources Reference Feng, S., Sekine, S., Pessino, V. et al. (2017...

...white arrowheads). It is necessary to use two different pairs of target sites for this strategy to work...rescued expression of GeneR.   Sometimes also referred to as DiO (Double-floxed inverse Orientation),...flip it to "off"). These Cre-off switches may be referred to as DO (Double-floxed Orientation) and could...the genOway press release for more information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling...

...academic labs, we have the capability to study many different mutants because the Caenorhabditis Genetics Center...domesticated housecat, and seeing how that makeup differs from that of wild cats with the goal of optimizing... space is the Hackerspace website. Markerspaces differ depending on the interests of their community, ...makerspace.ca, Victoria’s first hackerspace which now offers members and the public cheap access to personal...

...format for events and sessions in scientific conferences. They can result in lively discussions with both...the moderator is not a panelist and will play a different role so her ability to control the event is what...spontaneity so a moderator should be prepared with different questions than those on the prep calls, but often...stand and describe unique nonprofit models that differed from the companies represented on the panel. This...

...However, if your flow cytometer reads them out on different lasers, you should not need to compensate between... Guide Chart.  Pro tip! Some antibodies go by different names depending on your flow cytometer’s setup...compensation values (0-50). Inputting compensation values differs between machines or the program you’re using, but...look at the other channels you’re using first. A different fluorophore channel may be affecting your populations...

...means that any medium or washing buffer, such as PBS (Phosphate Buffered Saline), that comes in contact ...in a lab where multiple people are working on different things, this will help you avoid cross-contamination... Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling...

...or Förster] resonance energy transfer). FRET occurs when energy transfer from one FP (e.g. CFP) excites... acidic, or synaptic vesicles). Some FPs have different ex/em spectra (e.g. mKeima) or change fluorescent...Fluorescent Protein (GFP) Three Review Articles on the Different Types of FPs: Stepanenko et. al. (2008) “Fluorescent...

...the RPE65−/− dog, was characterized and found to suffer from early and severe visual impairment that faithfully...falling from grace, gene therapy is back, better and safer. It is now one of the most active and exciting areas...and the development of personalized medicine. References and further reading 1. FDA approves gene therapy.... PubMed PMID: 29983335. 9. Day TP1, Byrne LC, Schaffer DV, Flannery JG, 2014. Advances in AAV vector ...

...AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal cord...be identified by PCR later. The PNS presents a different problem: it’s spread throughout the body and is... and supplemental figure 8 in Chan et al).  References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered...selection yields AAV variants for widespread gene transfer to the adult brain. Nature biotechnology. 2016...

...park. Happy editing!   Resources and References References Yu, Y., Guo, Y., Tian, Qigi, Lan, Y., et...the break, potentially shuttling repair down a different pathway (Charpentier et al., Nat Com).   ...2014) 10.7554/eLife.04766 Moreno-Mateos, M. A., Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1...

...more flexible than Cas9/Cpf1. Some Cas13 enzymes prefer targets with a given single base protospacer flanking...are present in the gRNA spacer, as ADAR will preferentially edit an adenine if the template has a cytosine...that a S375A mutation in RESCUE (named RESCUE-S) offered ~76% on-target C-to-U editing while reducing off-target... Valley contributed to updating this article. References Cox, David B.T., et al. “RNA editing with CRISPR-Cas13...

...While I can’t make it “all better” for you, I can offer some tips and tactics that have worked for my friends...miserable at home or work, your relationships will suffer in both the short run and the long run and so will...alone, it should be obvious that your career will suffer. And, yes, your partner CAN take the kids to the... or her own dinner if you are going away to a conference. Really. No one will starve while you are away...

... optical pooled screens in human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin...editing efficiencies in a number of cell types at different genomic loci — in some cases, over 20-fold higher...prime editor. B, Prime editing efficiencies at different loci in U2OS cells. Image adapted from Yan et ...that an A122D mutation in the rhodopsin domain conferred several new improvements. The new variant has ...

...can be used with or as fluorophores in several different ways. Researchers have used oligonucleotides in...biosensors. Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied and used in...were born. Aptabeacons can be created in many different ways, but the general premise is that an aptamer...Aptamers can be selected for these small ligands. References 1. Ma, Hanhui, et al. “Multicolor CRISPR labeling...

... The entry clone and destination vector carry different antibiotic resistance markers (indicated here ...STEP 1: Generate an Entry Clone There are a few different ways to generate our desired entry clone - human...interest: Find Gateway cloning plasmids here! References 1. Chee JY, Chin CF (2015) Gateway Cloning Technology... Digest Video Check Out Our Molecular Biology Reference Pages  ...

...Addgenie Hannah Dotson.  Resources and references References Genome-scale CRISPR-Cas9 knockout and transcriptional...properly amplify.  Amplification protocols therefore differ from library to library. We strongly recommend ...Addgene’s pooled libraries use these protocols.  References McDade JR, Waxmonsky NC, Swanson LE, Fan M. ...

...immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, packaging...equally potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs...Sontheimer lab used NmeCas9 with AAV to edit two different disease-causing loci in mouse liver.  Two other...have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW, Mali P, Wu...

... studies.     Jennifer Tsang contributed to updating this article. References Abudayyeh OO, Gootenberg...variations of Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition...Zetsche et al., 2015 ). Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends...

...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi E,...RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using Cas7-11   Use ...fibrosis, which can be caused by thousands of different mutations, all found in the same gene. If we only...

...if your model is correct or your data favor a different model. Finding evidence for or against a model.../goal, an approach, anticipated outcomes, and preferably a description of the payoff if the aim is successful...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

...options in our CRISPR 101 series!  References and resources References  Anzalone, A. V., Koblan, L. W. &...plasmids that have been optimized for expression in different species and cell types including (but not limited...101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled Library...

...that “… the emergence of the video abstract genre offers a low-barrier opportunity for researchers to leverage...to measure the impact of their contribution. It offers a 5-step process and several examples for creating...professors at the University of Washington in St. Louis, offer guidance on how to make a low-cost whiteboard video...WeShareScience or email Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric...

...of conversations with people who work in many different scientific fields. They also host AMA forums where...team with a description of our AMA and Joanne’s preferred time to start answering questions. r/Science has...AMA is now a great resource for any scientists to refer to if they have questions about science careers,...future. The AMA was also cataloged for permanent reference by the Winnower. The only small downside to the...

...Vader in "Return of the Jedi" While Vader was referring to the evil side of a mystical "Force," this quote...cannot be ignored. They have the potential to interfere with the function of resident ER chaperones and...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...Kaposi’s sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent...

...of veracity of the conclusions drawn. The main difference with the PLOS policy is that access of this data...perhaps similarly impoverished lab) might apply different or novel tools and assumptions to reveal findings...geneticists may track the motion of multiple animals on different occasions via video, generating huge amounts of...responsible for storage.  If datasets are too large to transfer to a journal repository, they are likely too large...

...flanking a FLOXed neoR gene. pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR...Schofield, 2015; Moser and Hirsch, 2016; Epstein and Schaffer, 2017) Hopefully you’ve been able to check out...cohesin gene inactivation in human cancer.     References 1. Russell, David W., and Roll K. Hirata. "Human...27280971. 10. Epstein, Benjamin E., and David V. Schaffer. "Combining engineered nucleases with adeno-associated...

...nucleotide insertions, dele­tions or other mutations, referred to as ‘indels’. While this is convenient for generating... iPSCs (or correct them back to wild type), differentiate them into somatic cells affected by the disease...iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones! We ...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering...

...of interest supports a variety of processes in different cell types. One great way to do this is to use...SuperPLD module is optogenetically recruited by different CIB constructs to the plasma membrane or lysosomes...future updates to the collection! Resources and references Additional resources on the Addgene blog Viral....org Optogenetics Guide Optogenetics Plasmids References Aljiboury, A. A., Ingram, E., Krishnan, N., Ononiwu...

...to a mouse model of muscular dystrophy. Three different splice site donor and acceptor sequences were ...reconstituted after integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains...greater and more consistent transgene expression. References: Allocca M, Doria M, Petrillo M, Colella P, Garcia-Hoyos...retinal gene therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out ...

...insertions, like a fluorescent protein knock-in, suffer from low efficiency when a standard dsDNA repair... secondary structure. Today, multiple companies offer long single-stranded DNA production services or ...Quadros et al. attribute much of this variation to differences in guide RNA efficiency. For example, in a two-guide...Quadros for assistance with editing this blog post. References Yang, Hui, et al. "One-step generation of mice...

...directly into fertilized mouse eggs, circumventing the need for enrichment and selection offered by mouse ...into either the pronucleus or the cytoplasm of fertilized mouse eggs. Or, to avoid handling embryos ex ...therapeutic applications that better human health.     References Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib...

...nutrient-rich and also the best source for mycoplasma proliferation. Plus, it is added to the media after autoclaving... contact with a nutrient source, they start proliferating shortly. Once they are present in the laboratory...Bioluminescence assay kits are available from different biotech companies which are enzyme-based mycoplasma...and a Bachelor's degree in Biotechnology.       References Degeling MH, Maguire CA, Bovenberg MSS, Tannous...