... below lists plasmids that contain a worm ( C. elegans, C. briggsae , etc.) gene or sequence: ID Plasmid...Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses...to generate transgenic nematodes, including C. elegans and C. briggsae. Other related miniMos plasmids ...General Tools Fire Lab C. elegans Vector Kit - Andrew Fire Lab. A set of vectors for C. elegans research, ...distributes stocks of C. elegans. Silencing Genomes - Cold Spring Harbor's guide to RNAi in C. elegans WormAtlas...microscopy experiments. The C. elegans genome is well studied and hundreds of reporters have been developed and...Hubbard Lab. Plasmids for the temporal and spatial control of gene expression in C. elegans by combining expression...

...common solution is to keep your plasmid at -20°C or even at -80°C, in this case your preparation can be eluted...periods of time, especially at room temperature or at 4°C. Storing DNA: temperature and longevity You should... stable for years. Plasmid DNA can be stable at 4°C or even room temperature for a short period, and there...preventing the activity of DNase. While this is very important to preserve the stability of your DNA, it will...

...with C. elegans. Researchers commonly use rectal bacterial infection to induce tail swelling in C. elegans...also used a photoswitchable CDK5 in C. elegans. See figures 4B and C in Zhou et al for details on these...-ERK signaling in the hypodermis and gut cells of C. elegans causes the same phenotype. Activation of ... an ERK kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR...control of kinase activity can provide greater spatiotemporal resolution than pharmacological or genetic ...everything downstream of MEK1), bypassing the upstream portion of the pathway and focusing on the downstream steps...protein of interest. If you plan to try this, it’s important to check that the function of the protein and ...

...MMR activation.  The first way is to introduce a C/C mismatch at or within 6 base pairs of the edit site...-50% when activation of MMR is avoided Introduce C/C mismatches or silent changes in wobble codons to ...recombination genes at low temperatures (30-34˚C).  Shifting bacteria to 42˚C for 15 minutes quickly inactivates ...required for maintenance. A temperature shift to 42˚C not only allows for activation of the red genes needed...PubMed Central PMCID: PMC2790811. 5. Thomason, L. C., Sawitzke, J. A., Li, X., Costantino, N. and Court...system to modify your target DNA, you first electroporate a linear donor DNA substrate (either dsDNA or...Expression of lambda red recombination genes. Electroporation of substrate DNA and outgrowth of bacteria....

...Biddle, M. S., Alende, C., Fotouhi, M., Jones, C., Ayoubi, R., Southern, K., Laflamme, C., Virk, H., NeuroSGC...T. M., Brown, C., Bandrowski, A., Virk, H., Edwards, A. M., McPherson, P., & Laflamme, C. (2023). Scaling....154034.1 Laflamme, C., McKeever, P. M., Kumar, R., Schwartz, J., Kolahdouzan, M., Chen, C. X., You, Z., Benaliouad..., H. S., Biddle, M. S., Smallwood, D. T., Weston, C. A., Castells, E., Bowman, V. W., McCarthy, J., Amrani...Biddle, M. S., Wright, A. K. A., Sousa, J., Weston, C. A., Chachi, L., Roach, K. M., & Bradding, P. (2019... monoclonal antibodies. It is also important to seek out supportive data on candidate antibodies from ...t always indicate the presence of supportive data. To be supportive, the data must show that the antigen...

...CCL1 chemokine (C-C motif) ligand 1 I-309, P500, SCYA1, SISe, TCA3 CCL11 chemokine (C-C motif) ligand 11...chemokine (C-C motif) ligand 13 CKb10, MCP-4, MGC17134, NCC-1, NCC1, SCYA13, SCYL1 CCL14 chemokine (C-C motif...chemokine (C-C motif) ligand 19 CKb11, ELC, MGC34433, MIP-3b, MIP3B, SCYA19 CCL2 chemokine (C-C motif) ligand...chemokine (C-C motif) ligand 20 CKb4, LARC, MIP-3a, MIP3A, SCYA20, ST38 CCL21 chemokine (C-C motif) ligand...CCL24 chemokine (C-C motif) ligand 24 Ckb-6, MPIF-2, MPIF2, SCYA24 CCL25 chemokine (C-C motif) ligand 25...chemokine (C-C motif) ligand 26 IMAC, MGC126714, MIP-4a, MIP-4alpha, SCYA26, TSC-1 CCL27 chemokine (C-C motif...CCL28 chemokine (C-C motif) ligand 28 CCK1, MEC, MGC71902, SCYA28 CCL3 chemokine (C-C motif) ligand 3 ...

...example gene targeted for tagging with GFP at the C terminus, resulting in one edited allele, or gene ... gene of interest and decide which terminus, N or C, to tag and which short amino acid linker to use. ...study the protein or structure of interest. For a C-terminal tag, as shown in the example above, we inserted...precisely where we need it, in this example, at the C-terminus of the gene of interest. To get the molecular...tagging mammalian cell lines, Drosophila cells, and C. elegans. In addition to generating fluorescent protein...lines can be found in the Cell Catalog on our data portal. The plasmids should be functional in many different...and the donor plasmid into the cells we use electroporation, a technique that briefly destabilizes the ...

...recent models of C. elegans molecular biology are based on the standard worm genome. While C. elegans are ...list of active spaces near you.   Learning about C. elegans with the OpenWorm Project One such space ...project. The main goal of OpenWorm is to simulate C. elegans as a computer model. When I first heard about...deconstruct and understand this complexity using existing C. elegans mutants. For instance, we use a mutant known... funded by someone else (KittyBiome has been incorporated as a non-profit entity and so far most of their...

... (AacCas12b) exhibits optimal DNA cleavage at 48 °C, a temperature incompatible to genome editing in human...Bacillus hisashii, BhCas12b that is functional at 37 °C but preferentially nicks the non-target DNA strand...nicking Cas9. This base editor can be used for A•T to G•C edits in protoplasts or plants of rice and wheat. ...containing these base editors Spatially and temporally regulated AAV-based genome editing In 2016, Jonathan...AAV-based genome editing system is regulated not only temporally with doxycycline, but it can also be regulated...

...: 95 °C for 15 min, 35 cycles of (95 °C for 30 sec, 60 °C for 1 min, 72 °C for 1 min), and 72 °C for 10...following parameters: 37 °C for 30 min; 95 °C for 5 min and then ramp down to 25 °C at 5 °C/min. Dilute oligos...parameters: Cycles 1-20 (37 °C for 5 min, 20 °C for 5 min); Cycle 21 (80 °C for 20 min). These cycling ...Incubate at 30 - 37 °C for 24 - 72 hr. 30 °C may enhance genome editing efficiency, but 37 °C is acceptable....cells to incubate at 37 °C for 3 - 7 days and allow the clones to incubate at 37 °C for 7 - 14 days. Vary...thermocycler and run the following program: 65 °C for 6 min, 98 °C for 2 min to extract gDNA. Measure the DNA...clones. Run sample in thermocycler: 65 °C for 6 min and 98 °C for 2 min to extract gDNA. Screen each clone...

...Yamada, K., Kuznetsov, I. A., Rand, E. A., Ochs, C. C., Jaber, Z., Gardner, K. H., & Chow, B. Y. (2022...eLife.84364.  Meiring, J. C. M., Grigoriev, I., Nijenhuis, W., Kapitein, L. C., & Akhmanova, A. (2022)...., N., Ononiwu, F., Pal, D., Manikas, J., Taveras, C., Hall, N. A., Silva, J. D., Freshour, J., & Hehnly...j.ceb.2015.08.005.  Tei, R., Bagde, S. R., Fromme, J. C., & Baskin, J. M. (2023). Activity-based directed ...structure, separate chromosomes during mitosis, support transport of organelles and vesicles, and more. Although...microtubule-cutting sword Microtubules are one of the most important components of the cytoskeleton. They provide cells...demonstrated its use to probe cell mechanics, vesicle transport, and cell division, but given the long list of...

...Only about 10 amino acids can be deleted from the C-terminus and about 5 from the N-terminus, but this... standard practice to attach the GFP to the N- or C-terminus of another protein. This may, however, interfere...consensus sequence (MG-) and prenylation requires a C-terminal consensus sequence (-CaaX). Both modifications...when GFP is fused to these sequences. When N- and C-terminal fusions are not tolerated the alternative...the reasons that this works well is that the N- and C-terminus of GFP itself are relatively close, thereby...G-alpha subunit fusions that do not tolerate N- and C-terminal fusions (Adjobo-Hermans, 2011). Structural...that generates the chromophore (Tsien, 1998). Importantly, this reaction requires molecular oxygen and ...

.... A., Amrani, N., Peng, C., Kramme, C., Savic, N., Pacesa, M., Rodríguez, T. C., Stan, T., Tysinger, E...Dalvai, M., Loehr, J., Jacquet, K., Huard, C. C., Roques, C., Herst, P., Côté, J., & Doyon, Y. (2015)....Heussler, G. E., Hearne, C. C., Qu, J., Inclan, Y. F., Hawkins, J. S., Lu, C. H. S., Silvis, M. R., Harden...this strategy yields C to A base edits, whereas in mammalian cells, it produces C to G edits. Adenine ... Cas3 must be paired with the Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex...and Cas12a (also known as Cpf1) Cascade C RISPR- as sociated c omplex for a ntiviral de fense; a complex..., S., Bietz, A., Waldhauer, M. C., Börner, K., Fakhiri, J., Schmelas, C., Dietz, L., Grimm, D., Correia...

...El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi, R., Ledent... the UAS/reporter gene, the same bait and prey libraries can be screened using both N- and C-terminal ...organisms, including bacteria, alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the ...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...

...#834 - C-terminal GST for C elegans expression GFP Localization pcDNA3-EGFP - C-terminal...expression pET GFP - C-terminal GFP for bacterial expression pPD95_75 - C-terminal GFP ...transgenes Worm unc-54, variety of worm gene promoters C. elegans vector kit - Collection of...of plasmids from Dr. Andrew Fire L4440 - RNAi in C. elegans Dupuy Lab and Lehner Lab vectors...N-terminal Flag-HA for mammalian expression pNIC-CTHF - C-terminal TEV-His6-Flag for bacterial...Flag-TEV for bacterial expression pCS2FLAG - N or C-terminal 2xFLAG tag in pCS for a variety... including Xenopus HA Epitope Tag pKH3 - N or C-terminal 3xHA tag for mammalian expression...

...doi.org/10.3390/v12050513 Hsieh C-L, Goldsmith JA, Schaub JM, DiVenere AM, Kuo H-C, Javanmardi K, Le KC, Wrapp...plasmids used for this study at Addgene which include a C-terminal deletion in the spike protein to improve ...Zengel JR, Sathyan V, Nguyenla X, Van Dis E, Catamura C, Yamashiro LH,  Begeman A, Stark JC, Shon DJ, Fox ...Wrapp D, Lee AG, Liu Y, Chou C-W, Byrne PO, Hjorth CK, Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang...around the world, the spike protein has had many opportunities to evolve. Several variants are now circulating...different spike protein variants that can be incorporated into pseudoviruses. The lab used these pseudoviruses...Addgene to better understand which human genes are important for SARS-CoV-2 entry into human lung epithelial...

.... W., Déjean, V., Dodson, C. A., Wallace, M. I., Mackenzie, S. R., Timmel, C. R., & Hore, P. J. (2016)...Polali, S., Anderson, E. D., Bell, A. M., Tzouanas, C. N., Avants, B. W., & Robinson, J. T. (2019). Magnetic...doi.org/10.1021/jacs.3c01076.  Wheeler, M. A., Smith, C. J., Ottolini, M., Barker, B. S., Purohit, A. M., ...Beenhakker, M. P., Kucenas, S., Patel, M. K., Deppmann, C. D., & Güler, A. D. (2016). Genetically targeted magnetic.... Andrew and Maria were encouraged by previous reports that described magnetic responses of cryptochrome...reproduced their EGFP findings since the initial report in 2023.)     Figure 1: Fluorescence intensity...frequent collaborator and is happy to give advice and support for follow-up studies. “We don’t have a vision ...

...PMC3725456. Schwarz H, Zhang Y, Zhan C, Malm M, Field R, Turner R, Sellick C, Varley P, Rockberg J, Chotteau...aliquot of our recombinant antibodies is stored at 37 °C for a period of two weeks. After this time, quality...33151027; PMCID: PMC7665837. L'Abbé D, Bisson L, Gervais C, Grazzini E, Durocher Y. Transient Gene Expression...Sederstrom D, Steinmann D, Mozziconacci O, Schöneich C, Freund E, Ricci MS, Carpenter JF, Lengsfeld CS. Do...for further processing. It may be helpful to incorporate regular feeds during this incubation time to ...immunohistochemistry, and immunocytochemistry. It’s important to note that some antibodies are better suited...Chotteau V. Small-scale bioreactor supports high density HEK293 cell perfusion culture for the production of...

...information, you can fuse each FP fragment to the N- and C-terminal end of each PIP and empirically test which...R, Björklund M, Ahr D, Ahmed AW, Spinelli L, Brun C, Basler K, Merabet S (2018) Generation of a versatile.... eLife 7. https://doi.org/10.7554/elife.38853 Hu C-D, Chinenov Y, Kerppola TK (2002) Visualization of...doi.org/10.1101/pdb.prot076497 Kim Y, Jung JP, Pack C-G, Huh W-K (2018) Global analysis of protein homomerization...https://doi.org/10.1101/gr.231860.117 Kodama Y, Hu C-D (2012) Bimolecular fluorescence complementation ...conditions when compared to FRET. However, it’s important to note that BiFC is irreversible — FP fragments...interact or not? — FRET is preferable when the temporal dynamics of PPIs are the focus of study. Many...

...chain of BoNT/B into two fragments (BotNT-N and BotNT-C) and joined each fragment with one part of an established..., and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids at Addgene...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists...heavily on fluorescent protein genes. These optical reporters have limited deep-tissue performance, and sometimes...Mikhail Shapiro’s lab has developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression...