...screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but this tends to limit... (figure 4f) of the tTA inducer virus, but the desired sparsity of labeling is achieved with lose doses...-color labeling of neurons, because it doesn’t require the generation of a transgenic mouse. 2. Cell-... gene vectors for neurons, no transgenic mouse required (see figure 5 and supplemental figure 8 in Chan...

...binding-dependent expression of the HIS3 gene, which is required for histidine manufacture and therefore growth...of interest, or a defined cDNA library (e.g. the entire proteome of a particular cell type, or a subset...at the same time in order to try and access the entire interactome of a set of proteins at once.  However...gain access to the nucleus. To circumvent this requirement for transmembrane proteins, a split ubiquitin...

... Chen co-founded Addgene in 2004. They worked tirelessly to get Addgene from a concept to an up and running...previous years, this event was held at Executive Director Joanne Kamen’s home- a testament to the familial...it! 8. Organize If you’re doing an event that requires people to be in groups, organize your troops prior... this saying is especially true. If your event requires a lot of set up or cleanup, ask for volunteers...

...hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai virus requires stringent biosafety...to the process of collecting fibroblasts, which require a more invasive skin biopsy procedure. This becomes...PMC2917235. 11. Nakagawa, M., et al., Promotion of direct reprogramming by transformation-deficient Myc. ...

...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...single model. The first approach can lead to new directions, while the second approach could terminate with...think realistically about the time and resources required to feasibly accomplish the aims and clarify what... and Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia...

... the ideal codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...necessary for proper protein folding (Stadler and Fire, 2011). Therefore, codon optimization may disrupt...1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and its potential...doi.org/10.1016/s0165-0270(02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation...

...DNA strain which results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al., 2016). For TREE ...different reporter proteins. Protein translation requires the start codon of AUG immediately downstream ...

...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...translation of replication machinery proteins, or directly inhibit replication by binding to the origin of...sequences within the origin of replication and are required for replication in plasmids that contain them ...Kamruzzaman M, Shoma S, Thomas CM, Partridge SR, Iredell JR (2017) Plasmid interference for curing antibiotic...

...skill developed during PhD training. Creativity is required to develop new projects and test novel approaches... all the funding in the world? What if you could hire more people? What would you do with unlimited time...over half of which, when implemented, impacted the entire company. It is important to note that the person...product team is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism...

...Rett syndrome experience regression of previously acquired skills such as speech, coordination, purposeful...programs in biopharma. AveXis, co-founded by Kaspar, acquired by Novartis and rebranded to Novartis Gene Therapies...adeno-associated viral (AAV) vector. The editase repaired half of the mutant MECP2 RNA in each of three ...Samaco RC, Zoghbi HY (2011) Adult Neural Function Requires MeCP2. Science 333:186–186 . https://doi.org/10.1126...

... and therefore may need fewer resources for the entire translation process. For example, during high growth...inappropriate locations or fail to make it through the entire transcript resulting in the production of nonfunctional...that their codons are more appropriate for the desired expression host. This is known as “codon optimization...Addgene now have full sequence data, we recommend directly analyzing gene sequences for codon optimization...

...AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches Split...trans-splicing (62%) vector. 4. Sequential homology directed repairs Overview: The sequential homologous recombination... donor templates using two sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains...M, Colella P, Garcia-Hoyos M, Gibbs D, Kim SR, Maguire A, Rex TS, Di Vicino U, Cutillo L, Sparrow JR, ...

...be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs are also small...to avoid animal use by using semi-synthetic or entirely synthetic VL and VH sequences. Phage display also...for the scFv (Gaciarz and Ruddock., 2017), but requires an oxidizing environment to form. When expressed... age-related macular degeneration (AMD) since direction injection to the eye results in high drug concentrations...

...Ciuffi, 2012). One family of viruses, retroviridae, require integration for their life cycle to continue. They...IN (integrase). These viral packaging genes are required for the production of lentiviral particles.  The... keep in mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral... (2008). Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following...

...variety of behaviors, opening up new research directions that were inaccessible with earlier tools. They... in localization to the cell membrane and not a direct change in the fluorescent properties, pYtags are...variations and heat transfer at the subcellular level requires a probe with fast kinetics and high signal/noise...temperature indicator B-gTEMP designed to meet those requirements (Lu et al. 2022). By fusing two bright fluorescent...

...bypasses the 5′ mRNA cap required for eukaryotic systems and instead directly engages the ribosome for... protocol and are compatible with single-end or paired-end read flow cells. FREQ-Seq is an open source... The kit contains all the variant PUF segments required to bind any 8 base RNA sequence. The segments ...

...secondary antibodies altogether and multiplex with direct IF, using primary antibodies that are conjugated...However, the primary-secondary workflow (a.k.a. indirect IF) is more flexible and efficient, since you ...primaries for whichever targets you like. Plus, indirect IF amplifies the signal since multiple secondary...design. Like selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk...

... on as a dimer, depicted on the right, with the entire monomeric sequence duplicated in tandem, resulting... with large inserts (>7 kb)(Berg et al., 1989), direct repeats (Ribeiro et al., 2009), and at high copy...frequency plasmid recombination mediated by 28 bp direct repeats. Mol Biotechnol. 2008 Nov;40(3):252-60....Summers D. Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability...

...overnight.  In contrast, a semi-dry transfer only requires enough buffer to saturate the sandwich and typically...without any transfer buffer. These systems, however, require you to purchase expensive system-specific sandwiches...on the antibodies and labels being used and may require some trial and error (Yang et al., 2012).  Choosing... they provide qualitative measurements. If you require precise quantification of a protein, consider using...

... degree. I loved parasitology. However, when I enquired about roles in this field, most jobs needed at...understood why parasitology roles in the field require experience. Working in industry: Microbiology ...redundancies can be made within the company; new staff hired in anticipation of the contract suddenly have less...institutes. The lab at the University of Queensland required a scientist for a 6 month project to validate ...

... to stain all DNA. Some amount of expertise is required in this process because result interpretation ...serum to be used for subculturing. Avoid using expired products or reagents that have been stored for ...cross-contamination. Make sure you keep all the required equipment and reagents inside the laminar airflow... Biotechnology.       References Degeling MH, Maguire CA, Bovenberg MSS, Tannous BA (2012) Sensitive ...

...systems has led to extreme sequence divergence that requires a multi-pronged analysis to sort effectively. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named...select RNA transcripts, or be engineered to conduct direct RNA editing. There are four subtypes, A–D, with...surface-level function of each type could lead you to new directions in your research. We all have our preferences...

...University. Effective science communication is required to promote public support for research and to ...graduate student at Harvard University, former Co-director of Science in the News, and member of the SITN...her PhD from Harvard University, is a former Co-director of Science in the News, and is a member of the...

... rabies materials may be required for some countries and may not be required for others. You will be notified... notified if an export license is not required for your country after checkout, and the processing fee...

...after receiving the plasmid from Addgene you could directly start your digest, PCR, or transformation?  Great... we decided to include 10 plasmids that can be directly used in the research on the SARS-CoV-2. Nine of...experiments such as cloning, digestions, PCR etc. and/or directly long term storage. Every preparation is quality...

...The Developmental Studies Hybridoma Bank (DSHB), directed by David R. Soll, is a repository for monoclonal...most popular monoclonal antibodies include those directed against human transcription factors such as PAX7...to Our Guest Blogger!   David Soll has been the Director of the Developmental Studies Hybridoma Bank (DSHB...

... taking up a position on the Addgene Board of Directors. We're excited to watch both Addgene and Sample...How to Be A Good Manager from Addgene Executive Director Joanne Kamens Resources on the Addgene Website... Check Out the Other Members of our Board of Directors Become a Part of the Addgene Community ...

...every science field as we expand and encourage more direct conversations between scientists and non-scientists...process of starting a forum where non-scientists can directly approach scientists and doctors with their questions.... Use hashtags effectively. This would help you direct your content to the right audience. Avoid using...

...meteoric rise of CRISPR technology, the ability to direct enzymes – from nucleases to transcription factors...RNA. As a result, targeting the RNA of interest requires that the RNA be tagged. “This tool makes it possible...She also notes that for reasons that are not yet entirely clear some PUF proteins lose their solubility....

...information to make improvements. Members connect directly with one another – not just with the team leader...'t work to get people sharing.   Frequently ask direct questions for specific feedback in non-public settings...Scientists. Subscribe to the Addgene Blog to follow the entire series as it is published. For more reading try...