... the E1 gene from the packaging cell line is transferred into the AdV by recombination. It is therefore...for production of Gutless AdV can be found in reference 3. Production, amplification, and quality control...please see Addgene’s Adenoviral Guide webpage, and references 1, 2.*Pro-Tip*: once the correct recombinant ...specific for a viral DNA sequence. *Pro-Tip*: Ad5 reference material should be used as an internal control..., high-titer rAdV in your experiment. Enjoy! References 1. Luo, Jinyong, et al. "A protocol for rapid...

... 1-4 different gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different viral...embryos, and animals. Nanoblades can also carry different types of CRISPR cargo including RNPs for indel...can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote. As a proof-of-principle...versatile and accessible genome editing tool.   References Mangeot, Philippe E., et al. "Genome editing ...

...separately introduce each mutation independently into different mice, and then these strains should be crossed...When crossed to a mouse that is homozygous for a different mutation, the F1 offspring will be either hemizygous...include embryonic lethality, a short lifespan, or infertility and can arise due to copy number, expression ...transgene. These problems can be mitigated in different ways. Prior to engineering a transgenic strain...engineered mice. Best wishes on your experiment! References Becher, Burkhard, Ari Waisman, and Li-Fan Lu....

...activation by dCas9-CBP and the SAM system differ in target preference. Sci Rep 9: . https://doi.org/10.1038...programmable guide RNA. Gene activation by dCas9, also referred to as CRISPRa, was initially published in 2013...method, dCas9 is fused to CBP, an histone acetyltransferase domain capable of rearranging chromatin structure...experiment. Good luck with your experiment! References Bikard D, Jiang W, Samai P, Hochschild A, Zhang...

... Clinical Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells... synthetic promoters, the Roska lab used four different design strategies: Strategy #1 (ProA): Create... this library contained 226 AAVs, each with a different synthetic promoter driving expression of the same...was from a single class of cells  Of the four different promoter designs, promoters from strategy #4 had...ready-to-use AAV viral preps Browse all viral vectors References Juettner, J., Szabó, A., Gross-Scherf, B., Morikawa...

... we haven’t even imagined yet. References and Resources References Andrews NP, Boeckman JX, Manning ...consist of a mixture of antibodies recognizing many different epitopes. (Middle) Monoclonal antibodies produced...of Recombinant Antibodies Recombinant antibodies offer numerous advantages over polyclonal antibodies and..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P...

...updated by Jason Nasse in July 2023.  References and Resources References Chen X, Choo H, Huang X-P, Yang X...lead to such responses as decreased heart rate (Redfern et al., 1999) or altered neuron excitability (Zhu...to consider that the same receptor can elicit different responses depending on what cell type it is expressed....sciencemag.org/content/364/6436/eaav5282.long Redfern CH, Coward P, Degtyarev MY, Lee EK, Kwa AT, Hennighausen...

...difficult to study ncRNA, CRISPR systems offer a number of different options. By carefully considering your...without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are... you more about its function.  Resources and references Resources on addgene.org Addgene's CRISPR Guide... CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna...

...concentrating a viral stock. Different titering methods can reveal differences in physical versus functional...will be used in a couple of days, some researchers prefer to store freshly harvested virus at 4 °C instead...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate... Yarmush. "Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus...

... splitting transfected cells into 3 plates at different densities for selection). A 2% agarose solution...transfection into plates containing selection agent at different densities, then dispose of the plates that do ...freeze at least 2 ampules each of 4 clones with different levels of expression of our gene of interest. ...to rapidly isolate individual cell clones with different expression characteristics from a population of...cell clones picked using the agarose method do not differ grossly from those isolated using cloning rings...

...comes to fruit flies Drosophila start life as fertilized eggs that develop into embryos and then become...about one week from the initial deposition of the fertilized egg by its mother. Inside the pupal case, the... the heat. Isolating male and female flies Differentiating male and female flies is essential for setting...thorax to look like stubble (sb). For a better reference of various fly phenotypes, check out the Classical...latest information about gene function, while others offer fly stocks and other reagents. If you’re ever confused...

...create customized overhangs to assemble up to 6 different fragments in a prescribed order. These fragments...protocol is fairly flexible and adaptable to cloning different types of DNA, such as cDNA, PCR fragments, or ...process close to 100% efficient. Gateway cloning also offers a very high chance of success (>95% for a single...deposit your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly...Find vector backbones Read our molecular biology reference ...

...these vectors is controlled by a different promoter resulting in different levels of expression from each...functional, stable proteins. Protein expression refers to the second step of this process: the synthesis...well-defined media and can be easily adapted to fermentation allowing for large-scale, stable production ...labelled or modified amino acids that are useful in different downstream applications. Cell-free expression ...

...compare results across multiple targets because different CAPTURE cell lines will have varying levels of...lines Cell lines, primary cells Table 1: Key differences between original CAPTURE and CAPTURE 2.0 methods...that occur during development and cellular differentiation. Using a cell line model, the Xu Lab tracked...enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures chromatin's dynamic role....0 re-writes the script on gene regulation. References Han, J., Zhang, Z., & Wang, K. (2018). 3C and...

...randomized with a defined set of different trinucleotides (corresponding to different amino acids) in the regions...this, they injected the spike protein from two different coronaviruses into a llama over several days and...your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the protocol,.... Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput...

...library to identify RBD mutations which do not interfere with ACE2 binding, but impact recognition by specific...immunoprecipitate the entire complex in three different cell lines. The Gräslund lab plans to continue...10.1101/2021.01.30.428974   The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...of indels to measure time. These plasmids with different promoters driving expression of Cas9 were used...Janelia Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes ...

... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...media with supplements such as magnesium salts, buffering agents, and/or supplying additional carbon sources...

... alter the ability of one FP to nonradiatively transfer energy to the second FP, often decreasing the ...most cases, by using monomeric FPs or FPs from different families, which cannot form heterodimers (i.e....University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...avid long distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ...Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...

... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...backbones.  The backbone modifications listed can refer to changes made to the original backbone by the ...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...

...600 bp) and may take up precious space in viral transfer vectors with limited packaging capacity. Additionally...placement of the reporter gene). Addgene's collection offers a variety of plasmids to express two or more genes...Retroviral pEF1a-IRES-Neo IRES Mammalian MSCV-IRES-Luciferase IRES Retroviral pWPI IRES Lentiviral AmCyan-P2A-mCherry... new eBook! Our goal was to create a one-stop reference guide for plasmids. We've combined our Plasmids...solutions necessary for advancing the field. References: 1. Internal initiation of translation of eukaryotic...