... often weren’t easily accessible, due to a wide range of logistical barriers. Addgene worked to identify...availability vs accessibility issue was found in a less tangible form: knowledge and information about the materials...is achieved through interconnected incremental changes. It was only after creating a plasmid repository...learned about the importance of smaller, systemic changes, like using alt-text, captions, and inclusive design...

...reduce off-target effects of gene therapy, Feng Zhang’s lab has developed a system called DIRECTED (Delivery...Design). This modular approach combines an interchangeable set of fusogenic and targeting components to...where the fusion and targeting components are interchangeable. Image from Strebinger et al., 2023, reused...be tethered on the viral surface, expanding the range of targetable cell types. DIRECTED is compatible...

...skipped or whether selecting colonies and performing Sanger sequencing will suffice…and the answer is no. Due...screening - An excellent protocol paper by the Feng Zhang lab describing construction, lentiviral production...CRISPR/Cas Systems Forum- Moderated by the Feng Zhang lab, this is a useful place to ask experimental ...Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9 knockout and transcriptional...

...DNA 5' TTN Specific DSB in target DNA with 5' overhangs Cas13a (C2c2) 2x HEPN crRNA RNA 3' A, U, or... Cas13a targeting with a single crRNA The Feng Zhang lab used Cas13a from Leptotrichia shahii (LshCas13a...and cleavage through a series of conformational changes in the Cas13a molecule. Like Cas9, Cas13a tolerates...Ladha et al., 2020). Cas13a in mammalian cells The Zhang lab made the exciting discovery that some Cas13a...

...you to track a cell line’s age and monitor any changes. Your tracking system could be as simple as a piece... stocks to work from. The future Attitudes are changing but yet more is needed to convince people of some... to carry our independent quality control for a range of testing, from animal doping to forensics, DNA...Delivery Systems for Mammalian Cells Learn about the Dangers of Using GFP for Protein Localization Resources...

...10 closely related yeast strains. Adaptation to changing conditions - Organisms often need to express genes...conditions. With varied codon usage, an organism can change which proteins are highly expressed and which are...idea and has been used successfully in the past (Angov et al., 2008), but there are still many more features...bodies and is folding properly. References 1. Angov, Evelina, et al. "Heterologous protein expression...

... has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998). FLPe’s performance was further... single cell with all its synapses and how it rearranges these synapses in the living brain over time ...Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range Circuit Mapping.” Journal of Neuroscience 28(28)...Central PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart. 1998. “Improved Properties...

... Today in our transferable skills guide: Time Mangement for Scientists. Back when I used to do mammalian...immediate deadline - even the best of friends get hangry at some point. My cells, on the other hand, could...terrible handwriting, is copied below): Read new Zhang Lab Paper Find plasmids similar to those in the ...while I may have found splitting cells boring, hanging out in the tissue culture room also gave me time...

...institution, you can order plasmids covering a wide range of fields from Addgene. What you might not know ... Academy of Sciences 113.9 (2016): 2388-2393. Huang, Shijie, Xuechen Zhu, and Charles E. Melançon III...et al 2015[2]). The proteins encoded in this set range in size from 4 to 156 kDa, enabling the user to ...5373-5383. Walkiewicz, Katarzyna, et al. "Small changes in enzyme function can lead to surprisingly large...

...that certain polymerases such as Taq generate A-overhangs (a feature used in TA-cloning). Such non-complementary...near the 5’-end) and that also generates 4 bp overhangs. You can then use the Klenow fragment of DNA polymerase...polymerase I to generate blunt ends from these overhangs and ligate these blunt ends together in a standard...Xia, Yongzhen, et al. "New insights into the QuikChangeTM process guide the use of Phusion DNA polymerase...

...References Hao, Y., Yan, J., Fraser, C., Jiang, A., Anuganti, M., Zhang, R., Lloyd, K., Jardine, J., Coppola...reagents, researchers can measure this conformational change to investigate the role of specific integrins in...reagents to study the difference in cell morphology changes induced by integrins binding the RGD motif compared...

...caused by splicing defects, including ataxia telangiectasia, congenital disorder of glycosylation, and ...RM, Madhavan S, Pan X, Ran FA, Yan WX, Asokan A, Zhang F, Duan D, Gersbach CA. In vivo genome editing improves...Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe LH, Church GM, Wagers AJ. In vivo...Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. In vivo genome editing using Staphylococcus aureus...

...added, it binds tTA, resulting in a confirmation change that prevents binding to the promoter and turning...added, it binds cTA, resulting in a confirmation change that prevents binding to the promoter and turning...acid to Drosophila cells. Work by Wei et al. from Kang Shen’s lab has also shown that the Q system is also.... Wei, Xing, Christopher J. Potter, Liqin Luo & Kang Shen. “Controlling gene expression with the Q repressible...

...training in a globalized world where individuals can exchange ideas, network, collaborate, and innovate internationally...challenge. On top of that, in the midst of a rapidly changing, globalized environment re-emerging from international... University of Valencia): “IMP showed me a wide range of professional opportunities that are outside of... this conference, mentors and mentees met and exchanged impressions of the IMP experience. Conference ...

...-biophys-083012-130404 Zhang N, Zhang D, Chen SL, Gong B-Q, Guo Y, Xu L, Zhang X-N, Li J-F (2018) Engineering...flowering plant Arabidopsis thaliana (Li et al., 2013; Zhang et al., 2018) and the model bryophyte Physcomitrium...

...variety of backgrounds with technical expertise that ranges from astronomy to bioethics to chemical engineering...disease area. Depending on the organization, these changes to policy could shift how society interacts with...or essays. Note: the answer to this question may change over time and that’s perfectly normal!  Action ...what parts of your current job you want to keep or change and can inform your move into science policy. Read...

... Strebinger, D., Frangieh, C. J., Friedrich, M. J., Faure, G., Macrae, R. K., & Zhang, F. (2023). Cell...takes is a bit of chemistry magic to produce a wide range of useful molecules linked to that substrate. If... magnetic beads. In one innovative example, the Zhang lab used SNAP-tag to decorate synthetic viral particles...

...dispersion – standard deviation/error, variance, data ranges. how closely data points fall to each other (see...identifies a p value cut off. This method entails arranging all calculated p values from smallest to largest...– Tukey’s test, Newman-Keuls, Duncan’s multiple range test, and several others. Each test has different...negatives, moderate false positives. Duncan’s multiple range test – Most powerful. Low false negatives, high ...

...able to make the edits without any reagent-grade endangered Martian chicory root, but considering this is...you can better plan your transductions and use a range of reasonable MOIs. 2. Create a monoclonal Cas9-...lentiCas9-Blast (Plasmid #52962) was a gift from Feng Zhang and is described in Improved vectors and genome-...libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038...

... genes in mammalian cells (Bressan et al. 2017; Liang et al. 2017). Current methods of epitope tagging...Characterize clonal lines by ICC and PCR genotyping (Sanger sequencing); use primers flanking the tag sequence...References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso E, Matjusaitis M, Garcia-Diaz C, Blin C, Grant ...Rev Genet 32: 601-618. PubMed PMID: 9928493. 4. Liang X, Potter J, Kumar S, Ravinder N, Chesnut JD. 2017...