...Förstner, T. Keck,  K. Becker, et al. (2011). Nature Medicine, 18, 166–171. Pubmed. A. Ertürk, K. Becker, N. ...whole mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories...guest bloggers, Jae Lee and Pantelis Tsoulfas. Jae K. Lee is an assistant Professor at the University of...Leischner, A. Schierloh, N. Jährling, C.P. Mauch,  K. Deininger, et al. (2007). Nature Methods, 4, 331–...2012). Nature Protocols, 7(11), 1983–1995. Pubmed. K. Chung, J. Wallace, S.Y. Kim, S. Kalyanasundaram, ... 497, 332-337. Pubmed. R. Tomer, L. Ye, B. Hsueh, K. Deisseroth. (2014) Nat Protoc., 9, 1682-1697. Pubmed...of illumination (i.e. light sheet).  This is in contrast to confocal laser microscopy where the laser and...

...Chu AW-H, Zhou J, Lung DC, Kok K-H, To KK-W, Tsang OT-Y, Chan K-H, Yuen K-Y (2020) Comparative tropism,...Xiuyuan Ou, et al. that characterized the spike (S) protein of SARS-CoV-2 (Ou et al., 2020). The authors requested...Chen X, Chen J, Hu K, Jin Q, Wang J, Qian Z (2020) Characterization of spike glycoprotein of SARS-CoV-2 on...https://doi.org/10.1038/s41467-020-15562-9 Subbarao K, Roberts A (2006) Is there an ideal animal model for...fundamental part of the scientific research infrastructure. References Chu H, Chan JF-W, Yuen TT-T, ...

... J. P. K., Liu, M.-S., Hibshman, G. N., Dangerfield, T. L., Jung, K., McCool, R. S., Johnson, K. A., &...nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs to the NUC ...lobe—do not structurally resemble any other known proteins; they are completely unique to the CRISPR system... a single unpaired strand in the center of the protein. The repeat:anti-repeat duplex pokes out from the...RNA, confirming its sequence. This region of the protein varies between Cas9 enzymes from different species.... The stem loops bind along the outside of the protein, spanning both lobes. Stem loop 1, nestled between...shows the guide RNA stabilizing the DNA-binding protein portion of the CTD, ready to check for PAM sequencines...

... like chromoproteins, fluorescent proteins, and luminescent proteins. While fluorescent proteins and luminescence...interactions with cytosolic proteins. This step is important as non-specific binding to proteins can inhibit their...inquiries.  What is a chromoprotein and how is it different from fluorescent proteins? Before going into ...into the difference between chromoproteins and fluorescent proteins, it’s important to note that despite ...distinction by name, chromoproteins are actually a subset of the fluorescent protein family. What sets them...colors in ambient light. In contrast, the widely used green fluorescent protein (GFP) gives a bright green... history of fluorescent proteins in molecular biology research, chromoproteins have a shorter history....

...such as Rep or other initiator proteins, they do not require host proteins for replication and can maintain...elements, oris interact with various RNA polymerases and proteins from bacterial hosts. In a plasmid with...as recognition sites for replication initiation proteins. These iterons vary in their sequence and number... (ori1, ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various...has its own replication initiation factors and proteins independent of the host. Host range depends on...can replicate in different bacteria and express proteins within different host organisms. pBBR1-based plasmids...further to increase the range of hosts in which proteins can be expressed. For example, scientists can ...

... E. E., Yamada, K., Kuznetsov, I. A., Rand, E. A., Ochs, C. C., Jaber, Z., Gardner, K. H., & Chow, B. ...photoswitchable elements into many other types of proteins. Here, we’ll highlight a few recently-deposited...license. A naturally occurring microtubule-severing protein, Katanin is of course named for the samurai sword...optogenetic module to activate or recruit your protein of interest, maybe you want to inhibit it. Restricting...development and a knockout is not feasible or when the protein of interest supports a variety of processes in ...CIB1 clustering. Fuse either CRY2 or CIB1 to your protein of interest and express the other separately, then...generate CRY2-CIB1 oligomers that trap the fusion protein away from its usual location, preventing it from...

...interactions in the cell, including protein-protein, protein-DNA and protein-RNA interactions. The 1980s saw...Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein interactions...discoveries to create a new system for screening protein-protein interactions. In later years, improvements ... including protein expression and correct folding, post-translational modification, protein degradation... expression of target proteins in heterologous systems (e.g. mammalian proteins in S. cerevisiae). ...configuration of your fusion proteins physically blocks the binding sites for the protein partner or the UAS/reporter...C-terminal fusions of these proteins. This way both ‘ends’ of your protein are screened for binding. Using...

...with permission.   Mutations in the spike protein are one of the key drivers in determining whether...the current vaccines are effective or not. This protein both elicits an immune response as the target of... S, Wilkinson E, Naidoo Y, Karim F, Ganga Y, Khan K, Bernstein M, Balazs AB, Gosnell BI, Hanekom W, Moosa....2020.12.015 Garcia-Beltran WF, Lam EC, St. Denis K, Nitido AD, Garcia ZH, Hauser BM, Feldman J, Pavlovic.... In their assay, the pseudoviruses express a luciferase reporter which is used as a readout for infection...

... a complex of two proteins: the large (L) catalytic protein and its cofactor VP35. VP30 is a transcription...viral proteins (compared to viral infection). Therefore, the roles/functions of individual proteins can ... PubMed Central PMCID: PMC103829. 2. Conzelmann, K. K. "Reverse genetics of mononegavirales." Biology ...transcribed in the cell when the appropriate viral proteins are provided in trans (see Other Important Components... they do not encode all of the necessary viral proteins. Minigenome systems have been developed for many...genome and transcribes viral genes. Four viral proteins are required for viral genome replication and ...approach. First, this would prevent the study of the proteins independently from each other (see Uses of Minigenome...

...re-exposure (2, 3). More specifically, specialized Cas proteins snip the foreign DNA into small fragments approximately...paste them into the CRISPR sequence. Separate Cas proteins then express and process the CRISPR loci to generate...out these functions using slightly different Cas proteins and PAM sequences. A total of eight evolutionarily...us know.  References Ishino Y, Hideo, S, Makino K, Mitsuko A, Nakata A. J Bacteriol. 1987 Dec; 169 (... (2007). CRISPRdb. Date: 2013-09-18. Makarova K, Haft DH, Barrangou R, Brouns SJJ, Charpentier E, ... bacterial genome. Cas genes, which code for polymerases, nucleases, and helicases, were also disrupted...

...the folded protein (green region). Methanol dehydrates the protein, which can disrupt protein structure...and leaves many membrane-associated proteins in place. In contrast, Triton X-100 and Tween-20 are examples... (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction that covalently... polymerize and lose its ability to cross-link proteins. As such, you will get the best results if you...solvents can be quite harsh - lipids and soluble proteins can be lost during fixation with these chemicals...these chemicals can also alter the structure of proteins, which could disrupt your target epitope (Fig....antibody binding. Formaldehyde cross-links the protein (red lines), which maintains structural epitopes...

...detection because they produce very little or no viral protein. To solve this problem, two strategies have been... HIV so that infected cells will produce viral proteins and die, either through cytotoxicity or the immune...expression. In cell lines that can produce HIV-1 toxic proteins, CRISPR/Cas9 SAM caused apoptosis, indicating ...However, this approach could permit some viral protein production, so researchers are also interested ...antiviral therapies target HIV-1 at the level of protein structure, CRISPR gRNAs require DNA sequence-specific...Kaminski R, Fagan PR, Putatunda R, Young WB, Khalili K, Hu W. CRISPR/gRNA-directed synergistic activation...Tedaldi E, Napoli A, Zhang Y, Karn J, Hu W, Khalili K. Elimination of HIV-1 Genomes from Human T-lymphoid...

...specifically curated for protein expression in K. phaffii with special attention to protein modifications which...successfully for protein localization and transport studies in HEK cells, recombinant protein purification...used to optimize protein secretion and expression in the biotech workhorse yeast — K. phaffii (formerly...great option for those interested in recombinant protein purification and metabolic engineering. If instead...library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent option (Iverson...compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes...formerly known as Pichia pastoris). If your goal is protein isolation, then the MoClo Pichia Kit will get you...

...CRY2) and calcium and integrin-binding protein 1 (CIB1) proteins. The goal of both groups was to create...composed of two fusion proteins: 1) the genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1...form a functional protein due to being fused with high-affinity interacting proteins Coh2 and DocS, respectively...chimeric protein consisting of Cas9 and a photo-dissociable dimeric green fluorescent protein (pdDronpa... saCas9 protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly...photoswitchable proteins derived from the fungal photoreceptor, Vivid (VVD, N. crassa) (Kawano et al.,... Nihongaki et al., 2015 paCas9 Magnet proteins from N. crassa Split Cas9, Split Cas9 nickase, Split dCas9...

...fluorescent proteins in the nervous system. Livet J, Weissman TA, Kang H, Draft RW, Bennis RA, Sanes JR,...Brainbow-1.0 construct contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered...Brainbow-1.1 scales up the system to four fluorescent proteins, with recombination events specified by three ...principles remain the same. The first fluorescent protein in the construct, OFP (orange), is expressed when...invertible segments, each with two fluorescent protein coding sequences in opposite orientations. Cre-...Cre is removed from the system, the fluorescent protein whose coding sequence is closest to the promoter...fluorophores used are fluorescently distinct, their protein sequences are highly homologous, preventing design...

...few years, scientists have utilized CRISPR-Cas9 protein variants, Cas13, and Cas12a, to develop simple,...has adapted natural RNase activity of the Cas13 protein to develop and optimize the method termed Specific...endoribonuclease activity of the type III-A CRISPR-associated protein Csm6. RNA 22:318–329 . https://doi.org/10.1261/...S, Makarova KS, Semenova E, Minakhin L, Severinov K, Regev A, Lander ES, Koonin EV, Zhang F (2016) C2c2...Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara ...Semenova E, Minakhin L, Joung J, Konermann S, Severinov K, Zhang F, Koonin EV (2015) Discovery and Functional...sample is first amplified using RPA (recombinase polymerase amplification) or reverse transcriptase (RT)-...

...lymphocyte receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 ...170(5):956-972.e23 Screen identifies bromodomain protein ZMYND8 in chromatin recognition of transcription-associated...Epigenome CUT&RUN: An Improved Method for Studying Protein-DNA Interactions CRISPRainbow and Genome Visualization... NEAT1 RNA is essential for the structure of paraspeckles. Clemson CM, Hutchinson JN, Sara SA, Ensminger...

...mini counterpart label nearby proteins with biotin. The biotinylated proteins can then be isolated and identified...lysosomal-associated membrane protein 5 (LAMP5) — as well as cholinergic (choline acetyltransferase, ChAT) neurons. ...for Glypicans DNA editing with STITCHR Dynamic protein labeling with TurboCas Enhancer-AAV viral preps...our bacterial plasmid set for expressing tagged proteins, and stay tuned for mammalian expression and AAV...therapeutic potential. Our partners at the Institute for Protein Innovation (IPI) recently released a set of highly-specific...antibodies against three of the six human glypican proteins (GPC1, GPC3, and GPC4). All IPI antibodies undergo...structure. Glypicans are proteoglycans composed of a protein core, 2–4 glycosaminoglycans (GAG), and are (typically...

...many of the specifics - promoters used, precise protein sequences or domains, and methods of delivery -... at CRISPR. References Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V (2013) Plant genome editing ...10.1128/mmbr.67.1.16-37.2003 Khatodia S, Bhatotia K, Passricha N, Khurana SMP, Tuteja N (2016) The CRISPR...TTS, Sweeney CJ, Koman VB, Wong MH, Bohmert-Tatarev K, Snell KD, Seo JS, Chua N-H, Strano MS (2019) Chloroplast-selective...Wang C, Liu P, Lei C, Hao W, Gao Y, Liu Y-G, Zhao K (2016) Enhanced Rice Blast Resistance by CRISPR/Cas9...editing technologies especially in human cells. In contrast to homology directed repair, these methods do ...

...tissues, eliminating the need to deliver the Cas protein altogether. Platforming AAV Interest in interrogating...https://doi.org/10.1038/s41596-022-00700-y Hanlon, K. S., Kleinstiver, B. P., Garcia, S. P., Zaborowski...Chamberlain, J. S., Corey, D. P., Aryee, M. J., Joung, J. K., Breakefield, X. O., Maguire, C. A., & György, B....Danner-Bocks, S., Sanghal, T., Yadanar, L., Tian, R., Ma, K., Palop, J. J., & Kampmann, M. (2023). CRISPR screening...lentiviral vectors being packaged using VSV-G. In contrast, AAV can be packaged in a variety of capsids. ...