...they also uniformly stepped back to give a big picture perspective: Dr. Yamanaka talked about his clinical...trial patients with advanced macular degeneration; Laurenz Studer discussed his work with Parkinson's Disease...presenters, it was gratifying to see that Addgene has ensured that plasmid acquisition is not a rate-limiting...

...model organism for a few reasons: Affordable to culture and easy to work with Grows extremely fast (doubling...induced by removing methanol from the media. This feature makes the strain tractable for peroxisomal studies... for genome engineering. A unique evolutionary feature of K. phaffii, relative to S. cerevisiae, is that...

...receiving only 1 TF expression construct. Cells were cultured following a differentiation protocol before performing...so that a single TF was delivered to each cell, future applications could deliver multiple TFs to each...on the Addgene Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral Transduction...

...discuss those ideas and build from those.   Figure 1: Virtual lab meeting? No problem! You can still...final screen can also be saved and exported as a picture. I believe that showing something different from...document can be shared and serve as a springboard for future discussion. Collaboration and creativity are pivotal...

...got along well, suggest a coffee chat in the near future. Even just one more meeting (in person or virtually... but I’ll never forget the strawberries. Grand gestures are memorable! Attend events for one organization...defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is actually a map of Joanne...

...predict protein structures, then you might know a thing or two about computer modeling. Make sure to focus on...? Or perhaps you have presented in some public lectures? These transferable skills could be meaningful...

...see the Introduction to Fluorescence Microscopy lecture at iBiology. Commonly used filter sets & relevant...this filter set and have reported brightness measurements. Here, EGFP outperforms the improved folding...variants, presumably due to the lower growth temperature. This also suggests, however, that there is no...

...additional flexibility in pseudotyping.   Figure 1: Genome of a prototypical gamma-retrovirus.  ...nucleus through the nuclear pore complex. This feature may be useful for the transduction of terminally...-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to be descriptive...

...and you think there are four important times to measure that growth rate, you report the results of all...between hypothesis testing and generation, can be sure that results will be reported regardless of outcome...the journal and reviewers are incentivized to make sure that any possible null is really a true null and...

...processes directly affected by light. Long-term light exposure can also lead to cellular phototoxicity, and experimental...substrate like coelenterazine is supplied in the culture medium to allow RLuc8 to produce photons. The original...wavelength to specify which FP emission you’ll capture. Since the Nano-lantern system does not use external...

...results you've measured should change established theory. So, what can you do to ensure that your skills...our website and social channels to apply for our future events. In addition to this annual event, graduate...

...virulence. Scientists have studied how HGT occurs in nature and have learned how to introduce genetic materials...Transformation, transduction, and conjugation occur in nature as forms of HGT, but transfection is unique to ...material from the environment by bacterial cells. In nature, this genetic material often comes from adjacent...

...genes for “gas vesicles,” a gas-filled protein structure that water-bound photosynthetic organisms use ... check out this blog post. Bourdeau RW, et al., Nature. 2018. PubMed PMID: 29300010. miRNA reporter vectors... date, containing all 620 sequences of probable mature human miRNAs. Weiss’s collection consists of a...

...consolidate several steps into one. Soon you’ll figure out what prompts you need to consistently and accurately... accurately run a protocol.   Figure 1: Evolution of a Western blot protocol, from a full page ...experimental steps are worth doing, and doing well, so make sure you’re taking time to do them carefully and thoroughly...

...screening blog post for more information.   Figure 1: Result of a blue-white screen. Image from Stefan...more about colony PCR in this blog post.   Figure 2: The steps of colony PCR.   Sanger sequencing...in this case a plasmid. Sequencing is one of the surest ways to know that your insert is what you expect...

...won’t work if the fragment ends have secondary structure or too much sequence similarity. REPLACR-mutagenesis...expressing recombineering proteins Red/ET from a temperature-sensitive plasmid. Homologous recombination then...also well suited for insertions and deletions. The figure above shows guidelines for primer design; a desired...

...at all 1,500 or 1,600 transcription factors and figure out where they bind to the genome. We’ve been doing...other scientists quickly? Myers: We were hoping to figure out how to do this for many hundreds of transcription...as well. We would also like to put some social pressure on many other labs that may be doing something...

... known as the resting potential (Vmem), can be measured as the imbalance of electrical charge between ...of the pHLuorin fluorescent protein. This GEVI features increased brightness and improved plasma membrane...Levin, Director of the Allen Discovery Center. The future of this field has a high potential for the advancement...

...Find the CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed PMID: 30377362. Studying ...the cell membrane where cell voltage can then be measured. Archon1 is  bright, exhibits good localization...processes in vivo. Many photoactive proteins exist in nature, but developing them into optogenetic tools can...

...These workshops illustrate the open and modular nature of the workshop content: different parts could ...for securing funding to support this effort. The future of reproducibility workshops and lessons learned.... Be on the lookout for upcoming workshops at future conferences this year. Workshops are confirmed ...

...molecules of the RNA backbone to increase stability.  Nature has evolved ways to prevent degradation of important...and rendered useless after less than a minute of exposure to the appropriate light. The photoactivatable...to engage a target sequence. Upon a brief light exposure (several seconds) the gRNA is released and the...

... receive recognition for their efforts, inspire future research, and increase their impact. Open Access...review is the only way to rigorously determine or ensure a manuscript’s quality. But even journal-based ...  Stern BM, O’Shea EK (2019) A proposal for the future of scientific publishing in the life sciences. ...

..., with the expectation for major advances in agriculture, materials, chemicals, and medicine. As they’...and where all of it is likely to lead us in the future. Building circuits from CRISPRs Addgene: What...interactions. That’s relevant if you want to make sure you don’t target anywhere else, but for synthetic...

...communications and loves talking about science, nature, agriculture and bananas. You can follow her on Twitter...Plant Science today, Soapbox Science (hosted by Nature), and Addgene are open to guest contributions. ...

...increased pressure of the added liquid. The volume should be minimal in order to minimize the pressure into...mouse breeding colony) Environment Olfaction, temperature, humidity, noise level, circadian effects (inactive...

... Level 1 TUs into multigene constructs. Figure 2: The three assembly levels of MoClo. Image courtesy...modification of these chemical products. The flexible nature of MoClo allows users to test a variety of different....pone.0016765.  Hahn, F., Korolev, A., Sanjurjo Loures, L. et al. A modular cloning toolkit for genome...

...including propensity for off-targets, secondary structures, etc., as well as the presence of a Protospacer...one of several Cas9s that recognize NG PAMs. The Nureki lab generated SpCas9-NG, a rationally engineered...lab sought to address the PAM problem through structure-guided engineering of the PAM-interacting domain...

...Golden (Gate) pot (Pryor et al., 2022).     Figure 1: Workflow for high complexity Golden Gate Assemblies...cycling conditions (see Table 1).   Enzyme Temperature Incubation time/cycling Overhang length ...also optimized the thermocycler protocols and made sure that T4 ligase was the correct choice for the job...

...IgG2a antibody. When using a monoclonal antibody be sure to use a highly specific secondary antibody, rather... IgG2a>IgG2b>IgG1 (Manning 2012).      Figure 1: Broadly reactive versus subclass-specific antibodies...antibodies rather than generally reactive ones. And make sure to check for any reported cross-reactivity when ...

... have reached labs in over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image... the spike protein was developed based on 100 structure-guided spike designs and has improved protein ...Maynard JA, Finkelstein IJ, McLellan JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-...

...available, you now face the challenge of trying to figure out what system might work best for your particular...vector reseals and other incorrect products. This feature biases the reaction in favor of the desired assembly...which one works best for you. Happy cloning (and be sure to deposit your finished constructs at Addgene!)...

...depending on the size of the conference). This made sure I didn’t zone out and kept me engaged throughout...If you’re presenting a poster at a conference, be sure to practice beforehand. Generally, two types of ...be on their website or the program booklet. Make sure you’re familiar with the policies before posting...

...of antibodies as tools allowed us to detect and capture proteins of interest to better study them in vivo...tagged protein rather than at the C-terminus. These features made this tag easier to use and more flexible ...will remain an important tool for the foreseeable future. These newer findings simply add to our understanding...

...heterozygote into a homozygote and ensuring that all future offspring inherit the endonuclease gene. As we ...address many pressing global problems in health, agriculture, and conservation. But please be aware that there...biosafety officer should be familiar with appropriate measures. Barriers should be a component of all gene drive...

...vision in a canine model of childhood blindness. Nature Genetics 28, 92-95. PubMed PMID: 11326284. 4. Bennett...L., 2015. Gene Therapy returns to centre stage. Nature Vol 526, p.351. PubMed PMID: 26469046. 6. The Forever... Rev Virol. 2014 Nov;1(1):427-51. DOI: 10.1146/annurev-virology-031413-085355. PubMed PMID: 26958729.  ...

...range of attention spans, skill levels, and STEM exposure. If you expect a lot of kids, you’re going to ... we can make this fun activity relate to STEM.” Figure 1 shows the activities we had for my school’s Math...Perhaps you have young children and you want to make sure their STEM education is supplemented. A 2009 study...

...and Fultineer analyzed the gnomAD dataset, which captures a broader swath of human genetic diversity, to...Project: a global resource to map genomic diversity. Nature 604, 437–446 (2022). https://doi.org/10.1038/s41586...weaponization of genetics research by extremists. Nature 610, 444-447 (2022). doi: 10.1038/d41586-022-03252...

... mean a world of mental relaxation. And, I’m not sure I should put this in print, but I actually let my... my daughter skip school a couple of times. Be sure to add time with your family, for yourself and for...just “do science” – we are scientists. We have to figure out a way to keep being ourselves and to be happy...

...The detectable signal from this reporter can be measured and quantified to determine the presence and quantity...lateral flow strip format. They published this in Nature Biotechnology. Like the SHERLOCK detection system...2020) CRISPR–Cas12-based detection of SARS-CoV-2. Nature Biotechnology. https://doi.org/10.1038/s41587-020...

... that lack cookie monster are sterile due to a failure of primary spermatocytes to enter meiosis. Testis...Dominguez, Roberto, and Kenneth C. Holmes. "Actin structure and function." (2011). PubMed PMID: 21314430. ...segment number and polarity in Drosophila." Nature287.5785 (1980): 795-801. PubMed PMID: 9390559. Additional...

...Addgene. In vitro screening for the best gRNA Figure 1: Validation of double strand break (DSB) mediated...were fusing, you may have several mosaic mice. To figure out what alleles are present in these mice, you...favourite mammalian model organisms in the near future.  Many thanks to our guest blogger Samantha Young...

...behavioral tasks was an elevated plus maze that measures risk aversion/anxiety. The mouse can move freely..., learning, and memory. We’ll cover these in a  future blog post, so stay tuned!  References Alpár, ...rapid and long-lasting antidepressant effects." Nature communications 10.1 (2019): 223. PubMed PMID: 30644390...

...transduction was not detected. They also used an ELISA to measure binding of PHP.B to a soluble version of LY6A. ...to the central and peripheral nervous systems." Nature neuroscience 20.8 (2017): 1172. PubMed PMID: 28671695...for widespread gene transfer to the adult brain." Nature biotechnology 34.2 (2016): 204. PubMed PMID: 26829320...

..., the envelope gene, and a polyA tail.   Figure 1: Lentiviral production uses three plasmids: (...in Medical Virology (Li et a., 2018).   Figure 2: To pseudotype with the SARS-CoV-2 spike protein..., Bieniasz PD, Jäger U, Porter CD, Friedman T, McClure MO, Weiss RA (1997) Sensitization of rhabdo-, lenti...

...and S. cerevisiae (Tuller et. al., 2010). Figure 1: An overview of peptide synthesis. A ribosome...as Hoogsten base pairs and wobble pairs.  Figure 2: (A) The codon wheel depicts the 64 codons that...PM, Li W-H (1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and...

...than the other (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv...of panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of...Bates A, Power CA (2019) David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody ...

... dsDNA breaks: following a cut, the error-prone nature of non-homologous end joining (NHEJ) often leads...database, which relies on CAGE-seq to directly capture the mRNA cap, provides the most accurate mapping...examined the ability to use sequence-based and other features to nominate gRNAs that are likely to be active...

...pipeline processing (Nextflow), cloud infrastructure, and secure collaboration.  In that post, we talked...

...accompanied by new or refreshed figures made using BioRender.com. Figure 1: PRIME editing Please...

... significantly away from the functionality and features that previously made the platform a mainstay of...not delete @addgene on X. This will additionally ensure that the @addgene handle is always associated with...