...systematic studies of membrane protein interactions. Proc Natl Acad Sci U S A. 2004; 101(33):12242–12247. ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...

...Figure 2. PSAM4-based LGICs, their effect, and outcome in neurons. Please see the text or table 2 for details...receptors potently activated by an inert ligand. Proc. Natl. Acad. Sci. USA 104:5163–68. PMID: 17360345...with a specifically designed Gi-coupled receptor. Proc. Natl. Acad. Sci. USA 95:352–57. PMID: 9419379 Davies...G protein regulation of β cell function in vivo. Proc. Natl. Acad. Sci. USA 106:19197–202. PMID: 19858481... Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling...hM1Dq hM5Dq Human M3 muscarinic G αq CNO* Increase Ca 2+ Neuronal burst firing Armbruster et al, 2007 rM3D... details. Table 2. Descriptions of PSAMs and their activity in neurons PSAM Ion Pore Domain Ligand(s) ...

...genomics of biotechnologically important yeasts. Proc. Natl. Acad. Sci., 113, 9882-9887. DOI: 10.1073/...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...

... for real-time multicolor luminescence imaging.” Proc Natl Acad Sci U S A 112(14) (2015): 4352-6. PMID...improve the brightness of previous Nano-lanterns by 2-4 fold and permit five-color fluorescent imaging! ...activation, you can’t use fluorescence to monitor processes directly affected by light. Long-term light exposure...

...direct reprogramming by transformation-deficient Myc. Proc Natl Acad Sci U S A, 2010. 107(32): p. 14152-7. ...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ... to the process of collecting fibroblasts, which require a more invasive skin biopsy procedure. This becomes...measures, thereby increasing the costs of the procedure. Episomal reprogramming is an ideal and cost-effective...

...neurons with vesicular stomatitis virus vectors. Proc Natl Acad Sci USA 108, 15414–15419. PubMed PMID:...for retrograde transport has been found for AAV1, -2, -5, -7, -8 (Taymans et al., 2007), AAV1 (Hollis et... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310...reveal novel hippocampal CA2 circuits. Nat Neurosci 17(2), 269–279. PubMed PMID: 24336151. PubMed Central PMCID... Mechanisms of Motility and Regulation. Neuron 84(2), 292–309. PubMed PMID: 25374356. PubMed Central PMCID...transduction efficiency of recombinant AAV serotypes 1, 2, 5 and 8 in the rat nigrostriatal system. J Neurochem...analysis of adeno-associated viral vector serotypes 1, 2, 5, 7 and 8 in mouse brain. Hum Gene Ther 18, 195–...

...507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112... the genome of your favorite cells. Ma et al., Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7....precision in a native context. Chatterjee A et al. Proc Natl Acad Sci U S A. 2013 Jul 16;110(29):11803-8... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...

...immunodeficiency virus type 1 preintegration complexes.” Proc. Natl. Acad. Sci. U.S.A. 89 (1992) 6580–6584. PubMed...PubMed PMID: 8491198. PubMed Central PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human...

...immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl Acad Sci U S A. 2018 Feb 13. pii: 1722491115...JS, Legube G, Miller KM Genes Dev. 2015 Jan 15;29(2):197-211. DNA Cross-Bridging Shapes a Single Nucleus...would become busy and potentially detract from the process being depicted, we can’t forget that those other...plasmids through Addgene, don’t hesitate to start the process! Addgene depositors who study chromatin The PIs...

...with the endogenous tRNA-processing system. Xie K, Minkenberg B, Yang Y. Proc Natl Acad Sci U S A. 2015...3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication ...conserved cellular machinery to process gRNAs. Hijacking tRNA processing to make gRNAs In a recently published...in rice plants. Yang was intrigued by the tRNA processing machinery present in almost every living organism...gRNAs with any 5’ leading nucleotide could be processed, a distinct advantage over other methods that ...is its universality; robust and efficient tRNA processing is common to prokaryotes and eukaryotes. Though...

...extraction solution, runs the protocol, then uses around 2-4ul in each 50ul PCR reaction. The gDNA extracted ...facilitate the processing of the crRNA) is also expressed from a separate promoter. In the pre-processing form,...construct from our 'Split RNA' design, where the pre-processing form of crRNA is expressed and a tracrRNA (the...in cell and with Cas9 and tracrRNA, it will be processed (i.e., cleaved) into a shorter form that contains...our 'Chimeric RNA' design (meaning the mature processed form of crRNA is fused chimerically to a mature...

...protein expressed by recombinant vaccinia virus. Proc Natl Acad Sci U S A. 1991;88(20):8972-6. Moks T,...take care when using this method.      Fig 2:  A, The polyhistidine tag is a string of histidine...

...Illustration of SARS-CoV-2 from Philippa Steinberg for the IGI.   What SARS-CoV-2 plasmids from Ginkgo ... traditional, QC process. While this has worked well for the early sets of SARS-CoV-2 materials, it will...Bioworks to synthesize and share over 1,850 SARS-CoV-2 plasmids to help academic and industrial labs accelerate...request? Currently, plasmids containing all SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae ...bacterial expression systems, Ginkgo synthesized SARS-CoV-2 expression plasmids in a high copy backbone with a...high level expression and purification of SARS-CoV-2 proteins for downstream experiments and offer flexibility...optimized, full length and truncated versions of SARS-CoV-2 S (spike protein) cloned into a CMV driven expression...

...dual-transgenic recombinase-expressing mouse lines 2-9 and combinations of transgenic recombinase-expressing... present, the introns are excised during mRNA processing, producing a functional molecular tool (C,F)....Deisseroth Lab maintains a Standard Operating Procedure with general principles for working with INTRSECT...

...cancer. These viruses need to be handled in a BSL-2 (or BSL-2+) lab and require additional safety measures.... BSL-1. In BSL-2 labs, eye protection is required any time you have to work with BSL-2 materials, such...respiratory diseases in humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) ...recommended or required in BSL-1 and required in BSL-2 labs. When working with lentivirus, many organizations...sprayed into the air. For lentivirus and other BSL-2 materials, you’ll need to use aerosol-tight caps, ...percentage of the decontaminate to be used for BSL-2 waste, and of course, if you are working with a substance...plasmid (packaging plasmid). Viral production then proceeds by transfection of all three plasmids into a producer...

...neurons in the adult dentate gyrus and olfactory bulb. Proc Natl Acad Sci U S A. 2013 Mar 19;110(12):E1152-61... 2008 Dec; 18(6): 617–623. Pubmed PMID 19349161.  2. Deshpande A, Bergami M, Ghanem A, Conzelmann KK, ...foundational later on when we studied biological processes, like how our stem cells are housed in specific...G-deleted RABV can divide and replicate (since these processes are mediated by other viral proteins), but the...

...severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape ... SARS-CoV-2 infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface...Pooled library to study antibody escape in SARS-CoV-2 Recombinant antibodies for aminoacyl tRNA synthetases...viral preps Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz As multiple...viral spike protein.   Figure 1. (A) SARS-CoV-2 RBD DMS Library generation and sequencing. From Starr... binding ACE2 is the target of most anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes the...and a barcode, and is designed so that the SARS-CoV-2 spike RBD is displayed on the yeast cell’s surface...

...influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool,... (Phy) interaction 660 Cryptochromes Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein ...interactions based on Arabidopsis thaliana cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control...transcription effectors OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated... with single-spike temporal resolution. PLoS One. 2(3):e299. PMID 17375185 Kim B, Lin MZ. 2013. Optobiology...comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551 Saunders A, Johnson CA, Sabatini...

...the image can be acquired more quickly. 2-Photon Microscopy 2-Photon Microscopy attempts to solve two ...Fluorescence microscopy. Nature methods 2, 910-919 (2005). PubMed PMID: 16299476. 2. Conchello JA, Lichtman JW. Optical...enhance spatial contrast (i.e. optical sectioning (2) or super-resolution (3). Fluorescence microscopy...photobleaching and a reduction in signal intensity. 2-Photon microscopes restrict excitation (and bleaching...can be seen exiting through the tissue. Therefore, 2-photon allows for deeper imaging into thick tissues...imaging objective that collects fluorescence. Like 2-photon microscopy, only a single focal plane of the...relying on raster scanning as is done in confocal or 2-photon microscopes. Total Internal Reflection Microscopy...

...comprehensive SARS-CoV-2 virus-human interaction network in which we expressed 26 of the 29 SARS-CoV-2 viral proteins...all became acutely aware of the threat of SARS-CoV-2. Like many in the science community, we’ve been motivated...identifying these potential human helpers of SARS-CoV-2 (especially the ones that are druggable) is a key ...just at the beginning of understanding how SARS-CoV-2 interacts with human proteins. For example, we used... joining in this worldwide fight against SARS-CoV-2. As we’ve all been affected in various ways by the...details. Our set of constructs comprises all SARS-CoV-2 viral proteins except for Nsp3 and Nsp16, and includes...is going on there!).  Find the Krogan Lab SARS-CoV-2 plasmids (Note: The Nsp6 construct is undergoing QC...