...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...in education advocacy non-profits like Phipps Conservatory or the Brilliant Club and share your knowledge... photo competition. Or nominate yourself as an educator for a short, animated TED-Ed lesson. 6) Network... pros Where do you meet like-minded science communicators? Attend the annual flagship meetings of AAAS...bananas as a Marie Curie Fellow at The Sainsbury Laboratory. She is passionate about science communications...

.... The first was a new class of programmable riboregulators, called toehold switches, which allow for the...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...the technology through product development, the regulatory process, and manufacturing so that we can get...., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939...

...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...sterile truncated pipet tips. Indeed, an experienced operator can pick 48 clones from a 10 cm dish in 15- 20...YouTube video. Before returning the plate to the incubator, additional growth medium is added to about halfway...two 48-well plates of clones) which enables investigators to identify specific clones with desirable properties...Iris Lindberg is a professor of Neurobiology and Anatomy at the University of Maryland Medical School in...

...targeted tdTomato and pre-synaptic targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE...deployed around the world to combat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus...ready-to-use viral preps with more on the way. S5E2 regulatory element containing vectors developed in the Jordane...Flp-dependent DREADDs viral vectors. Both inhibitory and excitatory DREADDS from the Gether lab are now available...

...is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an engineer ...the toolkit. We have included a wider range of terminators than are commonly used as we have tested them...system. The paper also gives quantitative data for regulatory elements, which has not been available for plants...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have...start. They might want to use our promoters and terminators, but then they aren’t sure how to convert their...

...in a year-long biochemistry laboratory : Part II – research-based laboratory – a semester-long research...confidence in their research capabilities and laboratory and critical thinking skills (Knutson et al.,...improvisation and the latter making him a more effective motivator (“be like Tony Robbins!” he encourages faculty...research-based learning in a year-long biochemistry laboratory: Part I – guided inquiry – purification and characterization...

... are successful base editors in rice, tomato, wheat, and potato. Target-AID and rAPOBEC based CBEs are... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...Zenpei, et al. "Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion."...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...

...the cytoplasm. That is, unfortunately, some investigators realize too late that they have missed the impact...environment where it will localize [4]. Most investigators are surprised to learn that FPs are often poorly...overestimation of fusion protein activity. We encourage investigators to characterize FP-fusion proteins functionally...Einstein College of Medicine in the Department of Anatomy and Structural Biology. His interests include the...Medicine. Her doctoral research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent...

...upregulate an alternative pathway or induce other compensatory mechanisms. How can protein function be dissected...protein in Rosetta E. coli. New Lentiviral CRISPR activator and repressor plasmids from Scot Wolfe's lab. .... These include Tet-inducible CRISPR activators and repressor plasmids. (Kearns et al., Development. 2014...basic parts (promoters, UTRs, coding sequences, terminators, etc) into individual Level 0 plasmids, or choose...unit (Ex: a promoter, 5’UTR, coding region, and terminator). Next, up to six Level 1 modules can be similarly...in optimizing TALEN assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN...of the gene of interest, C-terminal tag, plant terminator, and plant resistance cassette) in pUC19 based...

...platforms to reliably detect nucleic acids at the atomolar level. For more information read Addgene's Blog...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...Cathepsin L Functionally Cleaves the Severe Acute Respiratory Syndrome Coronavirus Class I Fusion Protein Upstream...TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. . PCP4 - ...

...blocker isolated from a designed repeat protein combinatorial library by in vivo functional screen." Scientific...production of a downstream peptide, the transcription terminators in this series are just the tool you need. The...this portion of the BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP ...characterized in Cambray et al., 2013. These terminators can be used to precisely tune your polycistronic...Kit for some insights on its potential from its creators. We hope you’ll find these kits useful in your...

...list from your current P.I., lab manager, or administrator containing all of the materials that you’ve ...financial values. All too often, we see early investigators making unnecessary short term sacrifices that...solutions to reach your goals. For example, a junior investigator turned down a faculty position because none ...Happy Sciencing, Damien   Damien Wilpitz is a laboratory research management consultant and coach in Boston...research scientists create productive and manageable laboratory environments. His business skills enhance his...

...construct (about 8 in Brainbow-1.0 mice.) The combinatorial effect of multiple, independent recombination...woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), commonly used to increase transgene...plasmids References: Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous... Lgr5 stem cells. Snippert HJ, van der Flier LG, Sato T, van Es JH, van den Born M, Kroon-Veenboer C, ...PubMed. Multiplex cell and lineage tracking with combinatorial labels. Loulier K, Barry R, Mahou P, Le Franc...

... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively... Figure 6: Diagram of ovary and oviduct anatomy with the equipment used for i-GONAD. A micropipette...  Many thanks to Channabasavaiah B. Gurumurthy, Masato Ohtsuka, and Rolen M. Quadros for assistance with...Rolen M. Quadros, Channabasavaiah B Gurumurthy, and Masato Ohtsuka. “Easi-CRISPR for creating knock-in and...10.1038/nprot.2017.153 PubMed PMID: 29266098 Ohtsuka, Masato, et al. i-GONAD: a robust method for in situ germline...

...light within a specific range of wavelengths. Excitatory channels allow positively charged ions such as...combine excitation and inhibition by using both excitatory and inhibitory channels activated at different... the opsin.      Figure 2: Diagram of excitatory (ChR2) activation by light showing ions flowing... a single pathway to activate various protein regulators at different points in the pathway. Photoswitches...., Shane, J. C., McKnight, D. J., Yoshizawa, S., Kato, H. E., Ganguli, S., & Deisseroth, K. (2019). Cortical...

...in immune cells have identified many putative regulators of immunity. However, in vitro screens cannot...responses to cancer. The screen recovered known regulators, such as Ptpn2, Pdcd1, and Zc3h12a, demonstrating...Moreover, the screen identified putative novel regulators of CD8+ T cell responses in tumors, demonstrating...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...

...establish your own lab and get an independent investigator grant (e.g. an R01 from the NIH, in the United...can be found during conversations with senior investigators. Discussions can reveal mutual scientific interests...makes time for you, as needed, this is a strong indicator that this person cares about your science and ...manuscript reviews, etc. You'll also meet future collaborators and get access to unpublished information and...Einstein College of Medicine in the Department of Anatomy and Structural Biology, and now serves as the Director...

...customized to control transgene expression. Gene regulatory elements [promoters and enhancers, miRNA target...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. Many of...broad):  enhanced coverage for inhibitory versus excitatory neurons Mice, rats Yes (LY6A) Chan et al...AAV.CAP-B10 CNS: balanced coverage of inhibitory and excitatory neurons Mice, marmosets Unknown Brown et... frequently been used for screening candidate regulatory elements such as enhancers and promoters for ... its tropism is relatively more biased toward excitatory neurons than AAV-PHP.eB (Fig. 4c, Jang et al....., … Tasic, B. (2021). Enhancer viruses for combinatorial cell-subclass-specific labeling. Neuron,109(...