We narrowed to 867 results for: FER
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TypeBlog PostPublishedMay 15, 2018, 12:43 p.m.... 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...
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Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog PostPublishedMay 1, 2018, 1:57 p.m....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... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog PostPublishedJuly 23, 2020, 1:20 p.m....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... -
Split Fluorescent Proteins for Studying Protein-Protein Interactions
TypeBlog PostPublishedMay 3, 2022, 1:15 p.m....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...inducible genome editing (Feng Zhang Lab 2015) Reference Feng, S., Sekine, S., Pessino, V. et al. Improved... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....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... -
Experimenting in an Open Source lab: from CRISPR to Cats
TypeBlog PostPublishedJuly 23, 2015, 2:30 p.m....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... -
How-to: 5 Steps to a Great Panel Discussion
TypeBlog PostPublishedOct. 22, 2014, 1:57 p.m....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... -
Antibodies 101: Flow Compensation
TypeBlog PostPublishedMay 16, 2024, 1:15 p.m....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... -
10 Basic tips for mammalian cell culture
TypeBlog PostPublishedJuly 12, 2018, 1:09 p.m....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... -
Which Fluorescent Protein Should I Use?
TypeBlog PostPublishedMay 20, 2014, 2:06 p.m....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... -
CRISPR 101: Making a Knock-In Cell Line
TypeBlog PostPublishedDec. 22, 2022, 2:15 p.m....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... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....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... -
AAVs in Retinal Gene Therapy
TypeBlog PostPublishedJuly 31, 2018, 12:55 p.m....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 ... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....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... -
Negotiating Work and Life: How to Find the Joy
TypeBlog PostPublishedMarch 20, 2014, 3:04 p.m....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... -
Hot Plasmids Spring 2024
TypeBlog PostPublishedMay 23, 2024, 1:15 p.m.... 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 ... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog PostPublishedJuly 13, 2023, 1:15 p.m....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. ... -
Fluorescent Proteins 101: Aptamer Fluorophores
TypeBlog PostPublishedApril 11, 2017, 2:30 p.m....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... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog PostPublishedFeb. 17, 2022, 2 p.m....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... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog PostPublishedApril 16, 2020, 1 p.m.... 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...