...surgeries for viral injections can be found here [5, 6]. Opsin transgenic mice Transgenic mouse lines have...length of your projection, it may take as long as 6-8 weeks for sufficient opsin to reach the axon terminal...doi: 10.1002/0471142301.ns0120s59. PMID: 22470147.  6. Gore BB, et al. Manipulating gene expression in projection-specific...neurons disrupts reward consumption. Neuron. 2012;73(6):1184-94. doi: 10.1016/j.neuron.2012.02.016. PMID:... electrophysiology data too but that’s what collaborators are for). For example, if you study neural pathways...intended brain region and/or cell type (I’ll talk more about this in detail later). But what are the different...of your opsin, but keep you eyes open for posts about the next steps in the coming weeks. Expression of...

... CBEs are known to deaminate cytosine placed in 2-6 bp and 4-8 bp, respectively, of the protospacer. And... requirements Reference CBE SpCas9 rAPOBEC1 NGG 6 SpCas9 Target-AID (PmCDA1) NGG 5 SpCas9-VQR rAPOBEC1...biotechnology 35.5 (2017): 441. PubMed PMID: 28346401. 6. Zong, Yuan, et al. "Precise base editing in rice,...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...

...fluorescence-based ligand-binding assay. Scientific Reports, 6. https://doi.org/10.1038/srep25769 Grimm, J. B., &...Research. https://doi.org/10.1038/s41592-021-01338-6 Grimm, J. B., Muthusamy, A. K., Liang, Y., Brown, ...dynamics in mammalian cells. Biophysics Reports, 3(4–6), 64–72. https://doi.org/10.1007/s41048-017-0037-8...nanotoxicological studies. Environmental Science: Nano, 6(5), 1516–1526. https://doi.org/10.1039/c8en01381k ... A., Purkayastha, N., Flad, K., Schmidt, M. F., Tabor, A., Gmeiner, P., Hübner, H., & Weikert, D. (2020...

...writing Job changes can happen regularly (every 4-6 years is common), but this can also bring new opportunities—projects... company, likely will need to move around every 3-6 years—a short and exciting ride Complicated matrix...students Collaborations may be less “complicated” - for example, easy to initiate a collaboration without...up the pace Constant worry about grants and renewals What is different about non-academia? Scientific...make a decision about something like this without sufficient data? Starting to learn about the diverse options...with scientists just like you What is different about academia? Publishing in peer reviewed journals ...without having to worry about proprietary information Work hours are long, but there is more flexibility with...

...are then heat shocked to remove the SEC and after 5-6 days of growth, wild-type animals that have undergone...obtained the desired insertions on the first attempt for 6/7 genes. The data are clear - the SEC-based system...just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth... this species, namely, the robust HDR mentioned above, an accessible germline, and multiple selectable...modular system that requires very little hands-on labor and could facilitate the tagging of every protein...

...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence targeted...expression within 4 hours of blue light induction (6-fold faster than previously developed systems). The... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...neurons. Earlier this year, the Hegemann lab and collaborators report their discovery of a new family of seven...

...’re expressing 4 gRNAs, you’d use pX330A-1x4; for 6 gRNAs, you’d use pX330A-1x6. This customization means...specific genome editing.”  Nature Biotechnology 32(6) (2014): 569-576. PubMed PMID: 24770325. PubMed Central...PMC4090141. Find plasmids from this paper at Addgene. 6. Xing, Hui-Li, et al. “A CRISPR/Cas9 toolkit for multiplex...: CRISPR Optimized MAGE Recombineering." Sci Rep. 6:19452. PubMed PMID: 26797514 Find plasmids from this...multiplexing options, but also check out our blog post about multiplexing with Cpf1.   Figure 1: Multiplexing...assembly.  Unlike some of the plasmids described above, this vector does not contain Cas9, so you’ll need...similar to the Csy4 polycistronic system described above, except that they use an endogenous nuclease system...

...produce the glioblastoma model for this study (Ref 6)). In addition, it was exciting to see that the use... 543.7643 (2017): 113-117. PubMed PMID: 28225754. 6. Marumoto, Tomotoshi, et al. "Development of a novel...Cancer, Inflammation and Immunity and learn more about “... recent findings in basic immunology, cancer-immune...curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor Michael Karin from...This was followed by a talk from Professor Karin about alcohol and obesity as driving factors for chronic...got a chance to talk with Dr. Friedmann-Morvinski about tools used to analyse the tumor microenvironment...community, contact [email protected] to find out more about our free deposit and distribution service! References...

...experiments where I harvested a melanoma tumor from a C57BL/6 mouse and analyzed the tumor-infiltrating lymphocytes... gates around the populations of interest (Figure 6). A.                                                                                               ...                                                                                               B.      Figure 6: (A) A flow plot showing two stains in relation to...lymphocytes in a mouse tumor sample You made it! So much about the theoretical background. Let’s dive into the ...often choose a looser gate here. I am not worried about including irrelevant events in my analysis, as these...population, acquired from the previous gating pipeline, about 53% of my cells are dendritic cells and ~13% are...ve covered in this blog post. First, you learned about the types of scattering that occur when a cell is...

...cell type and another gene in a different cell type (6–8). Although it’s often very easy to get GAL4 UAS ... Microbe 22, 48-60 (2017). PubMed PMID: 28704652. 6. V. Rodríguez, D. Didiano, C. Desplan, Power tools... use the Gal4/UAS system, and you can read more about them in review articles (2–4). You can also take...

...was fortunate enough to be able to spend his last 6 months in graduate school focusing on concept development...from this decision, Mike and his co-founders raised $6 million. The biggest challenge during this time was...transitioned away from the bench and became all about finding the best product-market fit for this set...Notably, the team must have an upfront conversation about its goals and vision for the company; this will ...section! And read on for more great information about starting a biotech company. Suggested reading The...of Sample6) for taking the time to speak with us about the biotech startup process and work environment...

...Transduct Target Ther 6, 53 (2021). DOI: https://doi.org/10.1038/s41392-021-00487-6 Additional Resources...Viruses as Biological Tools blog can help you learn about the different types of viruses and pick one for ...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

...limited to a few strains, including 129 and C57BL/6, for which we have germline competent ESC lines. Second...Genome Editing. Current Protocols in Mouse Biology 6:39–66 . https://doi.org/10.1002/9780470942390.mo150178...Gene-Modified Mice using Staphylococcus aureus Cas9. Sci Rep 6: . https://doi.org/10.1038/srep32565 Resources on ... Delivery Methods Learn about Mouse Work on Exercise and Depression Read About Brainbow Resources on ...success is not always guaranteed. To learn more about this process check out our Mouse Modeling blog posts...by CRISPR. The sequence started clean and then, about 100 bps into the run, it got “dirty”, with multiple...makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies the target, the Cas9 endonuclease...

...., 2005 used a trans-splicing vector to deliver a 6 kb mini-dystrophin gene to a mouse model of muscular...Reassembly of Adeno-associated Virus. Molecular Therapy 18:6–8 . https://doi.org/10.1038/mt.2009.280 Hirsch ML,...genes to the retina by dual AAV vectors. EMBO Mol Med 6:194–211 . https://doi.org/10.1002/emmm.201302948 Trapani...first time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an AAV transfer...

... to reprogramming proteins for 8 hours a week for 6 weeks, and still required further culturing to generate...of Reprogramming Proteins." Cell Stem Cell 4, no. 6 (2009): 472-76. PubMed PMID:19481515. PubMed Central...cells by the piggyBac transposon." Nature Methods 6, no. 5 (2009): 363-69. PubMed PMID: 19337237. PubMed...PubMed PMID: 22258608. PubMed Central PMCID: 3652331 6. Shao, L., & Wu, W. (2010). Gene-delivery systems ...methods do you have experience using? Let us know about your iPSCs experiences in the comments below! References...Additional Resources on the Addgene Blog Learn about Methods to Reprogram Peripheral Blood Cells Check...

...place for drafting my stories. In total, I prepare 5-6 stories for an interview, since one story usually ...Addgene blog: Tips for writing a good cover letter 6 Steps to Submitting a Resume That Gets Seen Resume...you do to get ready for this interview? Maybe the above scenario sounds familiar to you. It was my experience...positions often required a one-hour presentation about a prior research project. In total, the interviews... find lists that have lots of general questions about job responsibilities, performance evaluations, professional...require. Frequently they will ask you to present about your prior research. This is a chance to highlight...

...developing one-on-one mentorship programs. Chapter 6 and Outro - Interview with high school student researchers...working in a lab for the first time, and thinking about the future. Many thanks to our guests,Olivia Ho-Shing...resources Use our Plasmids 101 eBook to teach students about basic plasmid biology Use or CRISPR 101 eBook to...

...regulatory functions.” Mol Microbiol. 1995 Mar;15(6):1031-7. PubMed PMID: 7623659. 3. Bernard, P., et .... 1992 Aug 5;226(3):735-45. PubMed PMID: 1324324. 6. Tam, JE. and Kline, BC. “Control of the ccd operon...in modern cloning, and why should you learn more about it? Read on to find out how ccdB can make your cloning...of this system for enhancing cloning efficiency about 20 years ago and developed cloning vectors to harness...

...expressing dCas9 as a means of modulating gene expression (6). Altering the length of gRNAs targeting different...PMID: 26436575. PubMed Central PMCID: PMC4747789. 6. Platt, Randall J., et al. "CRISPR-Cas9 knockin mice...of the Addgene Scientific Advisory Board. His laboratory at Columbia University is focused advancing scientific... Listen to Our Interview with Alex Chavez Learn about Sequencing Options for CRISPR Genotyping Cas9 Activators...

...01015-5 As for target selection, we usually pick 3-6 guides around the region to find the most efficient... Feng Zhang's Lab, has assembled a list of FAQs about using the lab's CRISPR technology. Genome..., February 2013 ), has assembled a list of FAQs about using the lab's CRISPR technology. Visit the Zhang...