We narrowed to 605 results for: RAN-1

...functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside the body...significant improvements in vision over a period of 1 to 3 years – the treatment appears to be both safe...research and science communication.     References 1. Gaspar, H. B., S. Howe, and A. J. Thrasher. "Gene...their mutations corrected, and the T-cells were transferred back into the infants. Almost all patients have...techniques may hold the key. In the simplest form, translating gene therapies from conceptual design to clinical...that carries a small, approximately 4.7 kb single stranded genome. AAV belongs to the family Parvoviridae...AAV vectors are also capable of infecting a wide range of cell types including but not limited to muscle...

..., check out Addgene's CRISPR Guide. Figure 1: Like this jar of jellybeans, a pooled CRISPR library... selecting a CRISPR library for your experiments. 1.  What species are your cells derived from? Currently...a different gene. Libraries may be available in a 1-plasmid system, in which Cas9 is included on the gRNA-containing... non-specific endonuclease (Cas9) and a single stranded guide RNA (gRNA). The ~20 nucleotide targeting...of wild-type Cas9 and a gRNA generates a double-strand break in the target DNA, which, when repaired through...scaled up for genome-wide screening due to the broad range of potential target sequences and ease of generating...heterogeneous population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within...

...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central... polymerase and transcription factors. Since the promoter region drives transcription of a target gene...repressor then prevents transcription from occurring, which means that transcription is now OFF.   We’ll...tetracycline-controlled transactivator (tTA). They fused TetR with the transcriptional activation domain VP16...

...label surface proteins with a membrane-impermeable biotin substrate (Figure 1). Biotinylated proteins were...adhesion molecules, transporters, ion channels, GPCRs, and more.  Figure 1: Workflow for cell-...564–571. https://doi.org/10.1038/s41586-022-05682-1 Zhang, Y., et al. (2025). Probing condensate microenvironments... and synthetic biology. Nature Communications, 15(1), 6563. https://doi.org/10.1038/s41467-024-50767-2...required, through mitochondrial transfer. In the two decades since this transfer was first proposed, researchers...manipulation transplantation (AMT), CRISPR-modified adipocytes (fat cells) are transplanted as a mass that...visible chromoproteins in a cell-free transcription-translation system (Figure 6). This approach eliminates...

... when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...expected in another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...application and available resources.     Figure 1: Table summarizing the IWGAV five proposed antibody...validation: A view from the mountains. New Biotechnol 45:1–8. https://doi.org/10.1016/j.nbt.2018.08.002  Uhlen...Interest (GOI), and transfect it into your favorite cell line. In parallel, you transfect cells with another...antigen - Recognition of an immunogen does not guarantee recognition of the target antigen. You want to...Antibody-independent methods may include analysis of transcriptomic or proteomic data from several different tissues...

...Zhang et al., 2015). In this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin...splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein domains that naturally...mutations with unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust...neurons from each class to fully characterize the transcriptome. The first data release for CeNGEN is planned... The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans. To study the role...using a split intein for refined spatiotemporal transgene control in Caenorhabditis elegans." Proceedings...

...and P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates are better suited for...non-selective plate. Most bacteria tested had 1 transfer in every ~100 to 1,000 colonies, but some bacteria...active for greater than 50 generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters...protocols can transfer CRISPRi into a single bacterial strain, such as a B. subtilis, or a narrow range of bacterial...can be introduced into a broader range of bacteria. Once transferred, Mobile-CRISPRi stably integrates...bacterial Tn7 transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes ...chromosomal copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during...

... (which range between 0 and 100) are then converted to a weight (which ranges between 0 and 1) so that...values (ranging from -40 bp to 40 bp) are also converted to a weight (which ranges between 0 and 1) (shown...distance) This gives each guide a score ranging between 0 and 1. The cutting frequency determination (CFD...summary per batch. Ranking guides for HDR in-depth In other tools, guides are ranked by specificity of ...genome by eye. We came up with a ranking formula that does just this: Rank = (compressed CFD score) * (gaussian...interactions (e.g., scores >80) are transformed to 100. This transformation (illustrated in the graph below...employs biological best practices. For example, it ranks guides by CFD score, and it mutates HDR donor templates...

...activating kinase 1 (MEK1) with either one or two phosphoserine residues cotranslationally inserted in their...directionally assembled into a Level 1 vector creating a single transcriptional unit (Ex: a promoter, 5’UTR, ...expression. Brown et al., Genesis 2014 Jan;52(1):39-48. APEX2 for proteomic mapping and electron...engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing the set of...comprised of three sets of cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly...coding region, and terminator). Next, up to six Level 1 modules can be similarly assembled into a Level 2 ...vectors to allow for additional iterations of Level 1 assembly if necessary. Combining multiple Level 2 ...

...Please see Figure 1 for commonly used markers for major mammalian organelles and transport pathways. It is...cell with the images from the Allen Cell Explorer (1). When studying the function of a protein or its role...Subcellular Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000....can be gained by understanding the location and transport of the protein within a living cell. Analyzing...Analyzing differences in protein localization and transport between healthy and diseased states can also provide...endoplasmid reticulum, modified in the golgi and then transported in vesicles via the secretory pathway to their...the end of their life cycle, proteins may be transported through the vesicles to the lysosome where they...

...homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in the PITCh vector ~20...editing possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted ...CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from ...genome edits. But have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated...primary methods for repairing DNA after a double-stranded break. HDR copies the sequence from a repair template...are chewed back to reveal homology, allowing the strands to anneal. DNA synthesis then fills in the gaps...5-25 bp) regions of homology flanking a double-stranded break yield precise deletion of the sequence between...

...biological questions. The two most common uses are: 1) to test the expression level in a specific system...brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g. TagRFP657, which...Photostability can be as short as 100ms (EBFP) or as long as 1 hour (mAmetrine1.2). However, for most FPs it is a...or Förster] resonance energy transfer). FRET occurs when energy transfer from one FP (e.g. CFP) excites...classified by the emission color (emission wavelength range) FPs are usually classified by emission color as...as outlined below (or emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP...nm Green 499 - 519 nm Yellow 524 - 538 nm Orange 559 - 572 nm Red 574 - 610 nm Far-Red 625 -...

... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...growth rate/transfection efficiency is not known, this is easily achieved by splitting transfected cells into...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day after...after transfection into plates containing selection agent at different densities, then dispose of the plates...under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to the gene of interest ...such as the Viaflo from Integra Biosciences) to transition from a 48-well cell plate to a 96-well assay ...

... products). I picked this post for a few reasons: 1) I think phage are super cool and have considered ...Connect the Research Community Grad School Advice Part 1: Picking a Lab and a Project That’s it for now. As...Editor's Choice Blog Posts As I’m sitting in the San Francisco International Airport listening to the Lion King...back from a very successful Outreach Trip in San Francisco and would like to give my thanks to all the Addgene...

...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome.... The activity of SpRY was validated in double stranded cleavage and base editing applications, with high...solution? Almost! Assuming a PAM of NRN, 50% of randomly selected genomic loci will have a PAM located ...

...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...Chapter 3: How all basic research ends up being translational. Ex: the discovery of GFP. Chapter 4: Goals ...

...backbones for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids ...rapidly in 2011, from zinc finger nucleases to transcription activator-like effector nucleases (TALENs). ...

... for less money. The key steps to colony PCR are: 1) design primers to detect the presence of your insert...primers. There are 3 strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers...previously described. When using insert-specific primers (1), positive clones (+) will give a band, while a negative...control. A good positive control is bacteria transformed with the same backbone plasmid. If this control...strain. A good negative control strain is an untransformed culture of the same strain of bacteria you used...Cloning Technique Learn to Perform a Bacterial Transformation Deposit Your Positive Clones with Us ...

...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6...PHP.B has a >40-fold increase in CNS transduction and transduces the majority of astrocytes and neurons... mice whose CNS was transduced by PHP.B but absent in mice who were not transduced by PHP.B. The Deverman...These results were surprising because in vivo transduction of AAV serotypes generally tracks between strains...diverse library of AAV9 mutants that had a small randomized peptide loop inserted in the AAV9 capsid. This... an enhanced version of PHP.B, shows similar transduction efficiency, but at a log lower dose than PHP.B...

...individual activities (1). Researchers also discovered the modular format of some transcriptional activators: that...requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References 1. Casadaban...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...flurry of discovery in the field of eukaryotic transcriptional activation and cell biology. During this period... that the DNA binding domain (DBD) and transcriptional activation (TA) domains retain their individual... regardless of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists...interactions. In later years, improvements to yeast transformation protocols also helped establish the yeast two-hybrid...

...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...Forster Resonance Energy Transfer (FRET). Through the non-radiative transfer for energy between neighboring...Whether BiFC constructs are expressed transiently by transfection or stably impacts the success of BiFC..., kinase signaling, integrin signaling, and transcription factor interactions (Kerppola, 2006). One of...interactions between basic leucine zipper and Rel transcription factors (Hu et al., 2002). This would be more...

...microscopic resolution for studying colloidal gold (1).  The method was based on using a thin plane (sheet...Siedentopf, and R,. Zsigmondy.  (1902) Ann. Phys.  315, 1 – 39.
 A. H. Voie, D. H. Burns, F. A. Spelman. (1993...microscopy/en_de/products/imaging-systems/lightsheet-z-1.html ...microscopy coupled with techniques that make tissues transparent are enabling microscopes to visualize the cellular...the cortico spinal tract, CST, spinal cord and cranial nerve afferents labelled with a AAV8 expressing...sectioning (OPFOS) (2). Arne Voie, David Burns and Francis Spelman focused a laser beam into a thin sheet ...of clear specimens and for in vivo imaging of transparent organisms. Advances in tissue clearing Similar...

...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production...only have to worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities...AAV and lentivirus requires three plasmids — a transfer plasmid containing your gene of interest plus ...in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus within...systems, it is standard to do only one passage (transfection). This differs from typical insect cell preparations...an additional couple of days, as it requires transduction of more cells, while AAV can be titrated following...

... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...non-selective anterograde transport to a lack of genetic control. ATLAS (anterograde transsynaptic label based on...of the color spectrum more powerful, the lab of Francois St-Pierre recently developed a pair of new yellow...in other plant species, thanks to the broad host range of TRV. Find TRV-TnpB plasmids here! Weiss, T.,...Viral delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis. Nature Plants...

...’re not one of these people yourself.  Fig 1: Quote from Michael Ende, courtesy of morefamousquotes.com... blog Management for ScientistsTransferable Skills Guide: LeadershipTransferable Skills Guide: Teamwork...the right time. Like many of the posts in our Transferable Skills series, this post is not meant to be ... that you could strengthen. The right set of transferable skills, coupled with your technical skills, ... can help, and if and when you find yourself transferring this skill to another job, it will be important...also aggravate long-term personal conflicts.  Transitioning a conflict into a collaboration may not be the...willing and able to solve, don’t take that skill for granted. When you go on interviews or are looking for a...

...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...possible, do at least two full length practices (~1 h) and get feedback. It’s also helpful to practice...aims do not need to be grant aims. Each aim could be the basis of a single grant. Either way, think realistically...talk could be your plans for your first NIH R01 grant proposal or a 10-year research plan. I suggest preparing...is a significant need for healthy livers for transplants. Growth of new livers in culture could resolve..., neither of these can substitute for a liver transplant. My lab will combine bioengineered scaffolds ... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...excellent choice for fusions with integral membrane proteins and membrane associated proteins (i.e. GPI-anchored...these compartments undergo significant post-translational modifications including glycosylation, disulfide... addition of N-glycans [9]. Both of these posttranslational modifications can disrupt FP folding rendering...including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...

...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the...would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...

...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging...requires more photons. For example, moving from transient overexpression of protein fusions to gene-edited... lipoic acid ligase, biotin ligase, phosphopantetheinyltransferase) – These enzymes catalyze the covalent... tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine)– Nonnatural amino acids can be ...improved brightness, photostability, and spectral range. Nevertheless, these fluorophores were designed ...

...my master’s, I worked in an academic lab where I ran a handful of assays over and over. Everyday was the...help you decide if a master’s might be for you. 1. Are you interested in master’s-level jobs? Ultimately...your work at conferences, help with manuscript and grant preparation, and lead your own research project....

...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike... and Screening via Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved...(MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand DNA (ssDNA...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

.... pH and DNases affect DNA stability Figure 1: So many choices! What to do? The main factors ...your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving...from the DNA. These apurinic sites if in double stranded DNA are repaired by base excision repair, but ...but can lead to mutations in single stranded DNA undergoing replication.   DNase on the other hand can digest...

... Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates transcription. The ... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...from one that represses transcription to one that activates transcription.However, this system lacks spatio-temporal...barbara_synbio) May 13, 2021 The original AraC transcriptional regulator Their system is based on the AraC...AraC protein that controls transcription from the PBAD promoter. “AraC is widely used in synthetic biology...It relies on the sugar arabinose to activate transcription by causing conformation changes in the AraC ...your gene of interest, all you have to do is co-transform pBLADEONLY. That way, a previously cloned gene...

... genetic tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site. GAL4 expressed...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with...visualize individual/non-overlapping cells. While transgenic animals are commonly used to specifically manipulate... to trigger recombination and expression of a transgene (Lin et al., 2018). However, this system requires...site to cleave and inactivate any read-through transcripts of the effector gene. Effector genes, any gene...across cell types and organisms by replacing the transcription factor binding site and effector in the construct... the SPARC system to use other experimental transcription activation systems, such as LexA/p65 or mCD8...

...our lives. Plasmid transformation into E. coli is a fairly inefficient process– just 1 out of 10,000 cells...down especially fast and plates should be used witin 1 month for optimal efficiency. Beware of satellite ...aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside...Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin...Polymyxin B polypeptide Alters outer membrane permeability Bactericidal 10-100 ug/mL Tetracycline tetracyclin...Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes...

...B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in..., with protocols using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template...sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand DNA incorporation (ssDI). "Easi-CRISPR...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...a DNA repair newbie - diagrams of resection and strand invasion are like a foreign language to me. But...supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "Mechanisms...

... similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already on a quest...nanobodies (Arbabi-Ghahroudi, 2017)!      Figure 1: Comparison of a heavy-chain only antibody and single-domain... tools for diverse applications. For example, RANbodies can be used as an alternative to conventional ...sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs in the clinic sdAbs have...also been some interesting work done developing transgenic mice that express camelid variable domains (aka...Llamas Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids...

...more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling as...Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...sequences to sequences in a database ExPASy translation tool: Translates a DNA sequence using all six open reading...Annotates features, BLASTs, aligns, digests, and translates a sequence.   NEBCutter2: For finding restriction...information, such as expression patterns, post-translational modifications and polymorphisms Codon optimization...

...organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...B) Strategy for leak-free transcriptional control. C) Iterative transduction workflow overview. Image ... by integrating a SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore...fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro, and they...

...cleavage. NgAgo takes a different approach, randomly removing 1-20 nucleotides from the cleavage site specified...About NgAgo User's First Impressions Here References 1. Gao, Feng, Xiao Z. Shen, Feng Jiang, Yongqiang Wu...in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast, RNA ...must be expressed from a plasmid or in vitro transcribed, and changes in gRNA secondary structure can ...removal RNA-guided CRISPR enzymes cause double-stranded breaks, with either blunt (Cas9) or staggered ...

...from distantly related organisms. In 2006, less than 1% of the genes deposited at Addgene were synthetic....synthetic biology is expanding rapidly.  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil ... synthesis companies often work within a narrow range for GC content. Rare codons also are not good predictors...sequences is Proteobacteria - the most frequent transfer of genes is from Proteobacteria to mammalian expression...natural gene sequence is, the less likely it is transferred into another organism in the lab. Predicting ...

...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ...expressing ECN is 109 cells/ml, which is within the range of  the ~1011 commensal bacteria per gram of stool...

...receptors. Biochimica et Biophysica Acta - Biomembranes, 1838(1), 15–33. https://doi.org/10.1016/j.bbamem...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes... dopamine D2/D3 receptors. Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020-78827-...for bacterial imaging . Microbiology Spectrum, 12(1), e0369023. https://doi.org/10.1128/spectrum.03690... of fluorescence: For applications that involve ranges of pH, and significant lengths of time, the stability...receptor conformation and signaling activity​​ (Sridharan et al., 2014). It's also crucial to evaluate how...the background. Dyes that can pass through cell membranes can prove useful for live-cell imaging. Experimental...

...this system.  Fig. 1: Cytidine deamination takes place on the free strand of DNA and converts a C...guide RNA (sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...changing its double-strand cut to a single-stranded nick. In order to produce a double stranded break, two sgRNAs... target DNA. The reverse transcriptase polymerizes DNA onto the target strand based on the sequence of...targeting dCas9 to transcriptional start sites was sufficient to repress transcription by blocking its initiation...approach is that two sgRNAs targeting opposite strands must be available in moderate proximity to each...

...  References and Resources References Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N.... Cell, 164(1), 18–28. https://doi.org/10.1016/j.cell.2015.12.041  Musunuru, K., Grandinette, S. A., Wang...may contribute to more in the future. Figure 1: A timeline of CRISPR base editing advances that led...the safety of their intervention, the team led by Kiran Musunuru and Rebecca Ahrens-Nicklas administered...disorder survive long enough to receive a liver transplant — creating intense time pressure for the research...al., 2017). These editors did not create double-stranded breaks in the DNA backbone, making them significantly... repaired with an A→G edit on the complementary strand. But the invention of base editing doesn’t necessarily...

...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9... the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987). The epitope (GKPIPNPLLGLDST) has...10.57733/addgene.l0ulu4. Hanke, T., Szawlowski, P., & Randall, R. E. (1992). Construction of solid matrix-antibody-antigen....org/10.1099/0022-1317-73-3-653. PMID 1372038. Randall, R. E., Young, D. F., Goswami, K. K., & Russell...functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They...

... RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated ...actually does, and neither plasmid is maintained (Figure 1).  The partitioning problem Plasmid incompatibility...Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs.77.1 Pubmed PMID: ... and inhibit its own transcription. As copy number increases, RepA transcription is repressed and replication...indirectly inhibit replication through inhibiting translation of replication machinery proteins, or directly... division. They don’t simply rely on chance (or random diffusion) to make this happen.  There are several...CBPs and NTPase to go around, the plasmids are randomly positioned, leading to plasmid loss (Diaz et al...

...sentences) What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your...Full "Applying for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need ...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly...emphatically deny that I have a magic formula to guarantee a reader the faculty job of his/her dreams. There...currency of scientific success is publications and grants. 2. Write succinct and easy to understand documents...years. You will discuss the focus of your first grant, how you will overcome certain obstacles, alternative...It's not about you and how you will finally be granted the faculty position you have craved since grad...

...other words, newborn poop) visible through their translucent bodies. This is the ideal stage to select for... transgenic flies...

...protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized ...several other constructs in plasmid form. Fig. 1: Summary of the approach and outcomes of the screen...voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...photoactivation of HcKCR1 in mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1...