We narrowed to 954 results for: COMP

...of the vector components are HIV-based? A7: No single plasmid contains all the components necessary to ...The components are divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs...decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells.  Q4: How...targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer...LTRs, PPT, RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more ...are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential for...

...have the full picture of the puzzle, the QC team compares it to the plasmid information or depositor-provided...depositor-provided sequences to confirm the correct plasmid components. For more information on this process, see our...small sample sizes, small regions of interest, or complex sequences such as G-C rich areas, secondary structures... NGS technology, due to the high-throughput compatibility. However, there are some instances where we ...thousands or even millions of base pairs. Several companies have worked on this technology, but Oxford Nanopore...sequence an entire plasmid in one read. Runs can be completed in anywhere from under an hour to 72 hours, with...data output. This method is great for resolving complex and repetitive sequences, but can be costly, low-throughput...

...  Here's what you'll find in this post:  New compact, efficient prime editors A versatile and precise...tape Teaching an old antibody new tricks   New compact, efficient prime editors By Susanna Stroik The ....), while PE6c and PE6d were most efficient at complex edits, such as long or highly structured templates...Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell, 186(18), 3983–4002...interchangeable set of fusogenic and targeting components to reprogram delivery vehicles like lentiviral...DIRECTED particles where the fusion and targeting components are interchangeable. Image from Strebinger et...the range of targetable cell types. DIRECTED is compatible with fusogens from different viral families and...

...intense amount of competition between microorganisms for available resources. This competition drives microbes...antibiotic compounds. In such a case, the microbe itself would be unharmed by the compound it produces...treat infections, we are treating microbes with compounds similar to those they have been battling for centuries...treatment regiment might wipe out bacteria that are completely susceptible to a drug while a full course of ...likely to develop over time. It isn’t feasible to completely eliminate the development of resistance; it’s...hygiene standards. Handwashing is an essential component of preventing infection, as is cleaning and disinfecting...

...the cell must have enough nutrients available to complete the entire round of replication. Initiation is...initiate replication. But pSC101 requires a third component: RepA, a protein encoded by pSC101 itself. RepA... are efficient for the host, as no additional components are necessary. Iterons are not the only tool ...Inc, also encoded by the ColIb-P9 plasmid, is complementary to part of the sequence forming this pseudoknot...restraining further replication.   Figure 3: Comparison of replication regulation mechanisms of stringently...replication. At high copy numbers, RepA proteins form complexes between iterons on different plasmids in a process...propagation by limiting their copy number and compensating for the additional metabolic cost imposed on...

...lab. Iron Chef-style competitions Every few months, we’ll have a cooking competition featuring a selected... eat together every day, sharing ideas for the company, but also talking about sports, movies, and just..., which is either an indication of the stiff competition or says something about my cooking.   This is... to organize. We’ll dedicate a post to these competitions in the future, and hopefully include some of...To-Do list long enough that there is no hope of accomplishing it all. How do we encourage people to take a... Hack Days, and we’ve seen great success at accomplishing “we-should-really-make-time-for-that” type projects...

...likely not a coincidence that the AAV genome is composed of single stranded DNA, which is particularly ... interest, ordering them from a gene synthesis company such as IDT or Genscript, and cloning them sequentially...infection with adeno-cre. AAV-based gene editing compared to CRISPR editing CRISPR, of course, has started...and virtually never exceeds 40%. While this is comparable or better than the efficiency of most current...systems. Ohmori et al 2017 use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template...researchers have devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied...2017 recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair...

...based on its library frequency before selection compared to its library frequency after selection. Key ...and easy technique that can be used to generate complex libraries of mutant plasmids ready for deep mutational...libraries, they all have inherent shortcomings compared to one pot saturation mutagenesis. Error-prone...vector with Gibson assembly. While this is a comprehensive mutagenesis method, a drawback to using Gibson...scanning libraries. PFunkel: Of prior existing comprehensive mutagenesis methods, PFunkel offers the best...second universal primer is used to synthesize a complementary mutant strand (step 2 and 3). Then the uracil-labeled...PCR-based approach for generating a customizable comprehensive mutagenesis library that’s ready to be tested...

...now available. However, it can be difficult to compare their performance, as most translation initiation...directly engages the ribosome for translational complex assembly. PhD student Dejan Gagoski used SITS ...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of proteins...-tagged human ORF clones to allow testing and comparison of different cell-free expression systems; the... construct barcoded, locus-specific libraries compatible with Illumina next generation sequencing in order...libraries using a 2-step PCR-based protocol and are compatible with single-end or paired-end read flow cells...21408-21413. PUF Assembly Kit PUF proteins are composed of repeat segments that can be engineered to bind...

... Now that you’ve completed a resource budget, and know how to decide which components are the most critical...an account of the material resources needed to accomplish your science, and can include: equipment, reagents...then start identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline...research success. If you can get away with using computer modeling and collaborations rather than using ... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...equipment. That’s why a resource budget can help you compare the monetary and empirical value that helps you...from academia altogether and instead started a company around his own research. He was clear on his science...

...binary systems. In recent years, a number of more complex systems have also been developed which don’t fit... Bujard, both the promoter and transactivator components of the system have been engineered and improved...revolutionized genetic studies in flies. An additional component, the GAL80 repressor, adds a repressible element...UAS-driven gene expression. To further increase the complexity of the system, researchers have used split GAL4... developed by Lai and Lee. LexA/lexAop is a complementary system to GAL4/UAS that functions in essentially... cells and ~24,000-fold in human HeLa cells as compared to basal expression. Co-expression of repressor...because it allows them to be used together for complex genetic analysis in Drosophila or for precise control...

...microenvironment, immobilizing the components and altering the composition and dynamics. The researchers demonstrated...are available, but red GECIs have been limited compared to other colors. Two recent developments add to...proteins, PinkyCaMP is the first based on mScarlet. Compared to RCaMP3 and jRGECO1a, PinkyCaMP had better brightness...to condensates, disrupting their dynamics and composition. Reproduced from Zhang et al. (2025) under CC... 2025). The team used AAV9 to deliver CRISPRa components to adipose cells to activate genes involved in... students to explore cell metabolism. Plasmid components of CRISPRkit are now available through Addgene... the Qi lab's website crisprkit.org to request complete kits or find protocols and other resources. Find...

...Burry 2011). Complex experiments Finally, additional controls may be required for more complex experiments...expected response and have a baseline with which to compare the experimental group.     Figure 1: A...western blot several times with the same samples and compare the results across runs. If the results vary considerably...will briefly touch on.  Western blotting When comparing samples in different lanes in a western blot, ... to the controls listed above, viability and compensation controls are recommended for flow cytometry....example, propidium iodide only stains dead cells). Compensation controls address the spectral overlap that occurs... occurs during multiplexing. Compensation controls are single stained samples for each fluorophore and...

...Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst...Serotypes Serotype refers to the different protein compositions of AAV capsids, which determine how a virus ...what works great in one system or cell type can completely fail in a different system (Issa et al., 2023...designing and testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two ...Taschenberger et al., 2017).     Figure 3: Comparison of two astrocyte selective promoters demonstrating...Konishi, Y., Hosokawa, Y., & Konishi, H. (2014). A comparative analysis of constitutive promoters located in...vector-mediated transduction of the corticospinal tract: Comparison of four promoters. Gene Therapy, 28(1–2), 56–...

...some clones being outcompeted by others in liquid culture, dilute and plate the complete library of transformants...faces as it approaches a host cell surface are complex polysaccharide structures, many of which are easily... DNA from host cell nucleases (4). To further complicate matters, even when sequenced, phage genomes tend... for cell-binding abilities (6). This can be accomplished using a restriction enzyme with a short recognition...involves forcing DNA through a small opening using compressed nitrogen gas; this shears DNA at random locations...your resultant plasmid libraries into chemically competent E. coli and analyze transformants for appropriately...transformation should be performed using a highly competent strain that you can use for long-term library ...

...reduced transformation and transfection efficiency compared to the monomeric version of the plasmid (Maucksch...multimers have additional origins of replication compared to monomers, they are also able to replicate at...bacterial cells with multimers grow more slowly compared to those containing the monomeric version of the...and more prone to being lost in bacterial cells compared to monomers, although this can be prevented via...cannot be used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services ...frequency of multimerization, and in some cases completely prevent the formation of plasmid multimers (Bedbrook...produced the highest yield of monomeric plasmid compared to DH5α. However, more research is needed to determine...

...to detect a single, specific protein within the complex mix derived from cells or tissues. Westerns are...sample per lane, onto the top of a resolving gel composed of the crosslinked polymer acrylamide. In addition...proteins are immobilized onto a membrane, usually composed of a nitrocellulose or polyvinylidene difluoride...membrane.  In a wet transfer system the sandwich is completely submerged in a tank filled with a transfer buffer...confirming that the protein is the expected size as compared to the protein standards.  Protein abundance In...loading control when assessing protein abundance or comparing samples across lanes.  For a typical loading control...present in all samples. Uneven sample loading or incomplete protein transfer causes inconsistent levels of...

...technology, and good computer skills will give you an advantage. Strong writing, computer skills (Word, Excel...you can fully investigate and solve a problem or complete a project, overcoming inevitable failures and ... Nature Photographer/Artist Ecologist     Computer and writing skills Writing a thesis or dissertation...which are useful to many employers. Increasingly, computer skills are highly desired, as roles are becoming...Will you be coordinating projects? Do you need computer programming skills? Other things to consider...as forensics currently have too many applicants compared to the number of available roles). Make your resume...

... interested in are compatible with your microscope optics, you will want to compare the excitation and...fluorescent proteins currently available. Choosing compatible fluorescent proteins To choose a set of fluorescent...scope of this post. Are your fluorescent proteins compatible with your microscope optics? To determine if ...is efficiently collected by the microscope. To compare the match between a fluorescent protein and a filter...here's a screenshot from the Chroma Spectra Viewer comparing a standard Cy3 or Rhodamine filter set (Chroma...channels used for more abundant proteins or marking compartments. I hope this sheds some light on multicolor ...

...Jurkat T cells Complexes with DNA; causes strand scissions 100-400 ug/mL *Not comprehensive.   ** In eukaryotes...cells. Of course, these mammalian vectors must be compatible with the cell type they are tranfected into –...cells. Before getting into the mammalian plasmid components, it may be useful to describe the means of introducing...called transfection. Transfection is somewhat comparable to bacterial transformation (the introduction...void. These ORIs, however, require additional components expressed in trans within the cell for effective...ORIs, respectively. The presence of these viral components greatly reduces the rate of plasmid dilution ...

...change. Because of this, ABEs are only weakly compatible with many Cas domains beyond SpCas9. In order...lab set out to evolve ABEs with an increased compatibility to Cas homologs. Evolving better base editors...and editing speed and consists of the following components: A plasmid in the host bacterium expressing ...phage mutation rate Phage infection results in a complete ABE protein. If the phage delivers an active variant...variants with much higher base editing activity compared to the original TadA-7.10 enzyme. After testing...which edits 590-fold faster than TadA-7.10. It is compatible with all Cas9 and Cas12 homologs that the researchers...adenine base editor with improved Cas domain compatibility and activity. Nature Biotechnology. https://...

...loss-of-function or neutral, as compared to wild type, would be extremely helpful as a complementary approach. Also, ...characterized? Some have developed exciting computational algorithms, such as SIFT or PolyPhen to approach... Genetic Pathway Reference Set. These Gateway-compatible entry clones are the product of many years of...cancer phenotypes. These assays were designed to complement the one assay per gene approach that has been...genes. These should be used to create ever more complete “look-up-tables” of mutant function that could...approach: Phenotypic Characterization of a Comprehensive Set of MAPK1/ERK2 Missense Mutants. Brenan L...

...clones for complete functional knockout of your target gene. Mismatch nuclease assays are completely “blind.... The drop-off probe is designed to perfectly complement the wild type sequence and is not capable of ...precision genome editing workflow. Digital PCR comparison to other validation methods As mentioned, these...96-well plate without prior quantification. Comparatively, mismatch assays require between 200 and 500... in a high background of wild type targets. Comparatively, detection limits of about 5% mutant targets...PMC3773023. 7. Vouillot, L., Thélie, A. & Pollet, N. Comparison of T7E1 and surveyor mismatch cleavage assays...

...this complex holds a pair of DNA ends together, forming a paired end complex The paired end complex then...then ligates compatible DNA ends together, thus repairing the break. This is a simplified, streamlined...processed. Processing occurs prior to ligation as incompatible DNA ends interfere with that step. Accordingly...pop-in” insertion) provided a template containing compatible overhangs is available. Cas9 can also be altered...Biochemistry and Biophysics and the Lineberger Comprehensive Cancer Center at UNC Chapel Hill.   References...

... Mediator or SAM. SAM is an engineered protein complex for the transcriptional activation of endogenous...endogenous genes and it consists of three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA... assemble a synthetic transcription activation complex consisting of multiple distinct effector domains...with a BRAF inhibitor. (The test was the perfect complement for one conducted earlier by the Zhang lab for...gain-of-function screening when combined with a compact pooled sgRNA library. Our SAM-mediated screens ...sam.genome-engineering.org/. Crystal structure of Cas9 in complex with guide RNA and target DNA. Nishimasu H, Ran...transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, ...

...becomes more of an apples-to-oranges comparison. Addgene: Can you compare the TALEN kit to CRISPR/Cas9? When...research the kit now makes possible, and how it compares to his CRISPR/Cas9 system. Addgene: For people...other cells. TALENs are engineered proteins that comprise a DNA binding domain and an endonuclease domain...especially well-suited for use in stem cells. (See the complete protocol at StemBook). Addgene: How are you now...genetic background and do an apples-to-apples comparison of all those mutations. If you make induced pluripotent...pluripotent stem (iPS) cells from patients and try to compare those lines, they may all have the mutations but...

...fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral vector ...forming a vector that could create replication competent virus. Even though this system was efficient at...encoding 1) the lentiviral vector genome which was composed of the wild-type 5’ and 3’ LTRs, the ψ sequence...potential risk for the generation of replication competent lentiviruses (RCL) especially if you are working...positive cells could make the virus replication competent. The 3rd generation of lentiviral vectors was...8 kb) Potential for generation of replication-competent lentivirus (RCL) Efficient gene transfer Potential...Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human tropism can still infect...

...done in reverse. Assuming you want to become competitive for a Group Leader position, what will you need...requires that all of the experiments have been completed. Therefore, to be ready to apply for a Group Leader... you get more publications, you will be more competitive for fellowships. If you explore your system more...Discussions can reveal mutual scientific interests, complementary perspectives, a shared communication style. ...job search. Therefore, you must balance these competing goals. Knowing when to follow a lead and when ...master. While the calculus of these decisions is complex, here are a few guidelines to help you evaluate... much new data, speculate on what you hope to accomplish and how your research fits in with the current...

...techniques differ in important ways, but can also complement one another by providing both broad and specific... predict off-target effects of sgRNAs using computational methods. They identify possible off-target sites...the clinic. NGS-based methods provide the most complete information profile regarding putative off-target... with in silico prediction to create a more comprehensive list of off-target editing events. Two of the...tissue from whole animal models, no exogenous component required (e.g. dsODN), doesn't require high read...Vouillot, Léna, Aurore Thélie, and Nicolas Pollet. "Comparison of T7E1 and surveyor mismatch cleavage assays...the limit of detection for Sanger sequencing: a comparison of methodologies for KRAS variant detection."...

... The Association for Creatine Deficiencies: Compensating for SLC6A8 mutations in Creatine Transporter...channels typically expressed on other cell types can compensate for or bypass the dysfunction of another solute...cells to identify any solute channels that can compensate for the defect caused by SLC6A8 deficiency. “...where one of the X chromosomes is partially or completely missing in women. Bicuspid aortic valve (an aortic...with an emphasis on oligomerization, signalling complex assembly, and localization to the nucleus,” says...at McGill University.  These protein-receptor complexes consist of many parts that work together on the...gene, which codes for G𝛃1 subunits of G protein complexes, lead to a rare type of developmental delay, and...

...minutes of inactivating the CASANOVA complex, fluorescent CRISPR complexes were observed binding telomeric ...using blue light (Bubeck et al., 2018). They accomplished this by inserting a photo-sensitive LOV2 domain...indicator podcast segment The neurons in the brain compute using electrical impulses to coordinate emotions...sensitivity, responds quickly, is photostable, and is compatible with optogenetic control. The researchers also... the “2in1” vector cloning system - a Gateway compatible approach to simultaneously clone two genes-of-interest...2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and Blatt, 2012). The 2in1 Plasmid...

...preserves RNA structure and naturally occurring complexes present in the cell, it can also introduce non-physiological...interest. See figure 1 below for details on the key components of RaPID. Ramanathan et al. also tweaked BirA... BirA* homolog found in B. subtilis. BASU was  compared head-to-head with the E. coli and BioID2 BirA*...streptavidin signal (~250-fold enrichment) when compared to a scrambled RNA motif control). BioID2 had ...with BASU still generated a >30-fold enrichment compared to E. coli’s BirA*. Faster labeling kinetics also...HHCS-associated sequences had lower IREB2 binding enrichment compared to wild-type sequences and that loss of binding...Russell, P., & Guttman, M. (2014). Methods for comprehensive experimental identification of RNA-protein interactions...

...-Vvd showed substantially improved activity as compared to other recombinase/photodimer pairs. Created.... https://pubmed.ncbi.nlm.nih.gov/24390935/ Hypercompact genome editor CRISPR-Casɸ By Melina Fan The ...toolbox with their discovery of CRISPR-Casɸ, a hypercompact genome editor from the Biggiephage clade of ...cells. Casɸ from Biggiephage can eliminate competing mobile genetic elements. Image by Basem Al-Shayeb...the Doudna Lab. Scientists looking for more compact genome editors due to delivery constraints and ...efficiency and reduced RNA-off target footprint compared to current N-terminal linked SaCas9 ABE variants...in screens without any reduction in performance compared to larger genome-wide libraries. Find the library...

...cells, and animals. Some of these viruses are completely inert, but others can cause diseases. Nervous...One such control is dividing up the essential components of a virus into different plasmids. For a retroviral... are not encoded in the transfer plasmid, the component that physically integrates into the host genome...genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection cycle... Figure 1: Separation of retroviral packaging components onto distinct plasmids for safety.    Viral...Viral cargo-specific safety   After replication competency, the second concern with virus safety is the ...materials, you’ll need to use aerosol-tight caps, compatible with your centrifugation speed, to cover the ...

... SCF E3 ubiquitin ligase complex. In the presence of auxin, this TIR1 complex binds to the degron sequence...depiction of chromatin within a nucleus -> DNA compaction around histones -> Examples of histone-modifying...included for visualization purposes, but the key component of the editing assemblies are sensor-actuators...The tool-kit consists of highly useful vectors compatible with Gibson and Saptrap cloning systems to generate...Resources page.  Plasmids for delivering CRISPRi components by a single AAV vector are now available from... developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target DNA without cleavage until it'...

... a Cambrian-like explosion of software tools accompanying the CRISPR revolution. (See omictools for even... a 96-well plate and comes out of it. Biotech companies are known to have tools that do this internally...guides and primers in batch, optimized for HDR. Compared to wealth of resources for guide design, there...batch analysis because of its ease-of-use and comprehensive reports. With CrispyCrunch, you get the all ... ranking formula that does just this: Rank = (compressed CFD score) * (gaussian normalized cut-to-insert...is within the coding region, the algorithm then compares the guide sequence to every possible 23bp sequence...Francisco. Before starting his career in software, he completed a MSc in Psychology at McMaster University in ...

...derived from AAV9, their efficacy is generally compared to the AAV9 serotype or an engineered precursor...doesn’t lead to enhanced transgene expression, compared to AAV9, in BALB/cJ mice but is more effective..."Mouse Strain-Specific Tropism?" column. For a complete list of mice strains with LY6A-dependent tropism... by the Gradinaru lab (Deverman et al., 2016). Compared to AAV9, it effectively transduces neurons and...toward astrocytes, but its neuron transduction encompasses more molecularly-defined neuronal subtypes and...astrocytes and neurons, with greatly increased efficacy compared to AAV9 (Hanlon et al., 2019). This capsid was...brain regions, with reduced off-target expression compared to AAV9 in infant rhesus macaques. The AAV.PAL...

... stranded DNA breaks that commonly result in a complete loss of function while nuclease-deactivated Cas9...have several limitations. CRISPRko results in a complete loss of function meaning that if an essential ... more hits than the SAM library approach in a comparison study screening for vemurafenib resistance in...(Konermann et al., 2015). Root and Doench also compared the Calabrese library to screens performed with...that ORF and CRISPRa libraries can be used in complement to study gene function. Find the Calabrese pooled...gene function while using fewer sgRNAs. These new compact libraries will allow scientists to conduct genome-wide...transcriptional activation by an engineered CRISPR-Cas9 complex." Nature517.7536 (2015): 583. PubMed PMID: 25494202...

...mutations in the genome, but they’re inefficient compared to CRISPR/Cas9 edits that rely on double strand...only occur through editing by a APOBEC-Cas9n-UGI complex. So how exactly does this work? Figure 1:...editing efficiency can be easily determined by comparing cells expressing only eGFP expression to cells...DNA editing efficiency of APOBEC-Cas9 editisome complexes in real-time (St. Martin et al., 2019). To create...DNA editing efficiency of APOBEC-Cas9 editisome complexes. The eGFP reporters rely on the correction of ...frequency of genomic base pair edits of interest compared to cells segregated solely by a reporter of transfection... single base editing by APOBEC-Cas9 editosome complexes. Scientific Reports 9: . https://doi.org/10.1038...

...Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using...structures of CRBNmidi alone (A) and in a ternary complex with the multiple myeloma drug mezigdomide and ...on its own, with drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as ...matured considerably faster, with expression levels comparable to GFP during early embryogenesis (Figure 3).... through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet), and the new mScarlet3...Merritt D.M., Pe K., Cesar M., Hobert O. (2024) Comparative analysis of new mScarlet-based red fluorescent...than FRET-based mechanosensors (~60 kDa) and are compatible with common microscopy methods. Plus, the luminescence...