... here.   Virus production HEKs are the go-to ‘packaging cells’ for viral production because they express...need only transfect the ‘helper’ plasmids (viral packaging machinery) along with the viral plasmid containing...the cargo of interest, and HEKs will produce and package viral particles. The viral particles are then filtered...

...around the world were closed and not receiving packages. So the entire team mobilized to figure out how...subsequent fluctuations in distribution. I don’t have space in this blog to call out every Addgene Team because...workforce for when we need to be working at full capacity to serve the scientific community. First, we applied...

...weight (53 kDa) when run on an SDS-PAGE gel. Cancer Impact p53 is the most frequently mutated gene (>50%) ...disorder known as Li-Fraumeni syndrome. Recently some impacts of p53 mutations have been characterized as gain-of-function...

... SARS-CoV-2 are the lentiviral production and packaging vectors VSV-G and psPAX2, which were used in a...Repositories around the world facilitate the rapid pace of SARS-CoV-2 research by removing logistic hurdles...and financial support to maintain repositories’ capacity for storing, validating, and distributing such...

...the human interactors we found in our map have an impact on virus replication. While we and our collaborators...one are in a pLVX vector background, so you can package them into lentiviral vectors if you need to. Additionally...get notified!) Attacking coronavirus, one FedEx package at a time Since the publication of our paper, our...

...individually overexpressed and available for lentiviral packaging and delivery to target cells as a pooled library... 2023). Fig. 1: Barcoded TF library was packaged in lentiviral particles and used to transduce ...so it is an invaluable tool to be used in this capacity. Similar to the Human TFome Library, the MORF ...

...keep viruses frozen during shipment was having an impact on the samples. We therefore devised a few experiments...solution thawed, as the CO2 was confined to the tube headspace when the samples were frozen. This was clear by...tubes prior to thaw released the CO2 from the headspace, thus mitigating the pH effects of dry ice storage...

...Biology and, on occasion, would meet in the same space where BosLab exists today. Back then, it consisted...Kendall square, but we saw the potential in the space and decided to reactivate it. We started out by ...projects at BosLab. Some are entrepreneurs using our space to perform early-stage experiments. For example,...

...Archives allots for 25 MB of space per user, but, if you need more space, you can sign up for a plan. ... by iGreenEarth AMINO is a very simple app that packs important biological information, amino acid abbreviations...

...Discrepancies in transduction efficiency can have a major impact on experimental design and interpretation of results...transduced by the vector. One method for minimizing the impact of this phenomenon is to use a retrograde recombinase-expressing...cell bodies of the projection neurons reside.   Impact for scientists While the mechanisms by which retrograde...

... by spaces. File names with more than one word should be named in the camelCaseFormat since spaces between...the basics. However, these notes take up a lot of space and don’t tell you why you did something or what...

...Usually Destroyed Yes, Cpf1 cleaves 5' of the protospacer Multiplexing CRISPR-Cas9 options before Cpf1...which are often more difficult to transfect or package into viral vectors. Cas9 multiplexing vectors are...bind and cleave the lentiviral RNA, preventing packaging of the virus.  Reversing the orientation of the...

... applied, but these methods can have a negative impact on cellular health. Another limitation of traditional... them capable of diffusing in the extracellular space; they also have low immunogenicity and cellular ...less likely to be sequestered in the extracellular space. In addition, rAAV2-retro’s altered amino acid sequence...diffuse long distances within the extracellular space along an axon, this does not seem to be the case...

...at some point, decide early on what to do in a namespace collision. It may even be worth it to have extra... you're solving, but consistency is key. Some namespace collisions you may encounter: vector, sequence...Use tried-and-true tools We use Python, Django, Apache, and MariaDB. Using tried-and-true tools means ...

... resulting in variable gene expression that can impact observed phenotypes. Factors such as the growth...medium, growth temperature, and growth rate can all impact plasmid copy number in a cell. One can attempt... from a plasmid to the genome without greatly impacting the level of gene expression. For example, the...

...repeats separated by an 8-bp long asymmetric core spacer sequence. The asymmetry in the core sequence gives... double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...been created, which contain a unique asymmetric spacer "NNNTANNN", where "N" indicates which bases may...

... and impact the field - its encouraged to dream big! It is crucial that the goal and the impact of the...research plan Most grant applications restrict the space available to describe the proposed work. Thus, every...

... arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected and harvested...original plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will ...integration events. In addition, promoter choice can also impact transgene expression. The most accurate method ...

.... 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 ...the Doudna Lab. Scientists looking for more compact genome editors due to delivery constraints and ...

... Viral Vector Packaging Service Viral Production Viral Production Jump...cells using the 2nd generation psPAX2 and pMD2.G packaging system developed by the Trono lab . Cell culture...

...a 10 cm dish in 15- 20 minutes- a far more rapid pace than using cloning rings. A picture of a 60 mm dish...the plates that do not appear to be yielding well-spaced clones. For rapidly growing cells we sometimes ...the use of an electronic pipettor with variable spacing (such as the Viaflo from Integra Biosciences) to...

...Clutter not only makes it difficult to clean your workspace properly, but can also disrupt the laminar air...materials into the hood. This will give you enough space to move around freely without causing spills. Also... than 80-85% confluent because cells like their space. If they reach confluence, they stop growing (contact...

...are sharing stories of our repository’s positive impact on science and scientists around the globe. Our...CRISPR plasmids to labs in ~100 countries.  The impact Addgene’s CRISPR collection contained 197 plasmids...citations.  CRISPR is a story that centers around the impact of science, but our data makes it clear that it...

...will likely be spending a few hours on your feet. Pack water and snacks to keep you energized throughout...session, bring extra clips to hold up your poster, and pack a few business cards to tack up next to your poster...hearing the highlights so that you can appreciate the impact of the project as a whole. In addition to time ...

...other people to do that too. We don’t have the capacity to send tools out to people ourselves, but we’...committees, and how the culture of the university impacts the number of women reaching senior positions. ... those amazing students that want to have a big impact on the world and tell them to work in plant science...

...sequences, the amino acid sequences RSIAT and RPACKIPNDLKQKVMNH have been used successfully (Kerppola, 2006...expressed transiently by transfection or stably impacts the success of BiFC experiments. In either case...//doi.org/10.1101/pdb.prot076497 Kim Y, Jung JP, Pack C-G, Huh W-K (2018) Global analysis of protein homomerization...

..., including the E1 region). The space in the viral particle’s capacity that is made available by this ...genome, and adenoviral vectors all have a genomic capacity that is much higher than other viral vectors (...

...antibody (HCab), which are unique to camels, llamas, alpacas and other camelids. In terms of structure, HCabs...have done the hard work for you by immunizing an alpaca, isolating HCab antibodies from its serum, and ...for generating nanobodies are outlined below; no alpaca required. Decide Which Nanobody To Use: The table...

...interested in? Math, sarcasm, romance, and spacecraft! Image courtesy of xkcd.com Science communication... lectures, presentations, etc… Low (except for space) Real-time audience engagement and response. High...fear. Can be discouraging to build audience. Need space & higher entry barrier for audience versus passive...

...top):  A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins...INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and technical considerations...INTEGRATE system follows a simple workflow: Clone spacer(s) into the CRISPR array Clone desired DNA cargo...