...editing window, for example all C’s will edit to T’s. Thus, it’s important that there are no “bystander ... single base pair edit. (Cytosine deaminases edit C-to-T and adenosine deaminases edit A-to-G.) In short...takes place on the free strand of DNA and converts a C to U without producing a double strand break. ... particular gene isn’t feasible. For example, temporary repression or knockdown allow you to study essential...

...10.1126/science.1206593 Powers S, Miranda C, Dennys-Rivers C, Huffenberger A, Braun L, Rinaldi F, Wein....2020.107878 Sinnett SE, Hector RD, Gadalla KKE, Heindel C, Chen D, Zaric V, Bailey MES, Cobb SR, Gray SJ (2017... More recently, the Bird lab found that only a portion of the MECP2 protein is necessary to rescue the...Neurological Disorder by Programmable RNA Editing. Cell Reports 32:107878 . https://doi.org/10.1016/j.celrep.2020.107878...

... by isolating a calcium-binding protein, troponin C (TnC), from muscle cells. They identified a toadfish...calcium indicators can be used in mouse, Drosophila, C. elegans, and zebrafish – essentially all of the standard...said. “We know CREB is really important, but where is it really important and when? That is not known.”...calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes of these Twitch...

... GCR, GR, GRL nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor) NR3C2 Plasmids... MR, NR3C2VIT nuclear receptor subfamily 3, group C, member 2 NR4A1 Plasmids NR4A1, GFRP1, HMR, N10, NAK... RZR-GAMMA, RZRG, TOR RAR-related orphan receptor C RXRA Plasmids RXRA, NR2B1 retinoid X receptor, alpha...transcription factors modulated by ligands. Two of the important regions of nuclear receptors are: 1) a ligand ...

... Cre: Cre recombinase is split in half into N and C terminal Cre fragments (NCre and CCre, respectively...Bacterial Ekker 47551 pDD104 (Peft-3::Cre) Cre eft-3 C. elegans Goldstein 48201 CAG-GFP-IRES-CRE Cre and ...system CAG Mammalian Pelczar 51268 pCAG-Co-InCreC C-terminal component of the Co-InCre system CAG Mammalian...GFP (CRE-DOG) EF-1 alpha AAV Cepko 69571 pAAV-EF1a-C-CreintG Cre recombinase dependent on GFP (CRE-DOG)...dependent on GFP (CRE-DOG) CAG Mammalian Cepko 69573 pCAG-C-CreintG Cre recombinase dependent on GFP (CRE-DOG)...-Cre Cre CMV RABV Jessell 73717 pMLS328 Cre eft-3 C. elegans Jorgensen 75267 pAAV-CIBN-CreC photo-activatable...split-Cre CMV Mammalian Hirrlinger 106369 pCMV-Tag3B-CCre C-terminal split-Cre CMV Mammalian Hirrlinger 106370...

...nitrogen or a dry ice/ethanol bath, and store at -80° C. Once thawed, the AAV aliquot should be used quickly...PubMed Central PMCID: PMC190186. 9. Grieger, Joshua C., and Richard J. Samulski. "Packaging capacity of ...Axonal Transport It has been demonstrated that certain serotypes can undergo axoplasmic transport. These...of neurons (retrograde transport) or towards the synapse (anterograde transport), which is a possible ...retrograde and anterograde transport, but they also have similar transport characteristics (velocities...strong CNS tropism (4). If axonal transport is not desirable, transport can be reduced by injecting a smaller...and using AAVs have in common? The three most important items when looking for a house and choosing an...

...mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments in mammalian...can form it’s chromophore at a more comfortable 37°C (5). Dendra2 photoconverts from green to red fluorescence... pH stability, chromophore maturation rate at 37 °C, turnover rate, and the level of background fluorescence...: 1035-1040. PubMed PMID:18724362. 8.Stiel, Andre C., et al. "Generation of monomeric reversibly switchable...impossible to determine specific protein turnover or temporal expression using standard FP-chimeric proteins...second. The first successful irreversible PA-FP reported was PA-GFP. PA-GFP was derived from Jellyfish ...localization, or leading to protein aggregation. Another important parameter to consider is the contrast ratio- a...

...Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) ...Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung... https://doi.org/10.1126/science.aaq0179 Myhrvold C, Freije CA, Gootenberg JS, Abudayyeh OO, Metsky HC... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg...

...create solution C. And if a conflict resolver does their job well, not only will solution C be better than...many sources of conflict are substantive (“Person C told Person A one thing and inadvertently told Person... is customer support, but there are eight other teams who still retain a customer support element amongst... your technical skills, can open new career opportunities for you, if you can identify them in yourself...explaining my approach to conflict resolution, it’s important to recognize that there are different types of...or goal. Resolving these types of conflicts is important for the individuals involved, and also for the...responsibilities. Knowing which aspects of customer support each team “owns” is critical for day-to-day operations...

...LRRK2 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40376 pBACgus/LRRK2 1867-2141 C-His...2161 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40378 pBACgus/LRRK2 1867-2176 C-His...2161 C-His_T2031E LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40381 pBACgus/LRRK2 C-His_G2019S...2161 C-His_G2019S LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40383 pBACgus/LRRK2 C-His_C2024S...mCherry-MAPTau-C-10 MAPT mCherry CMV Parkinson's, FTD Michael Davidson 55311 mTagBFP2-MAPTau-C-10 MAPT mTagBFP2... mKO2-MAPTau-C-10 MAPT mKO2 CMV Parkinson's, FTD Michael Davidson 58112 tdTomato-MAPTau-C-10 MAPT tdTomato... CMV Parkinson's Mark Cookson 13314 pCMVTNT PINK1 C-myc PINK1 Myc CMV Parkinson's Mark Cookson 13315 pcDNA-DEST53...

...constructs 69570 and 69571 together encode the N and C terminal segments respectively of a GFP dependent ...-N-CretrcintG Connie Cepko 69571 AAV1 pAAV-EF1a-C-CreintG Connie Cepko 75469 AAV1 pAAV-EF1a-Flp-DOG-NW.... The advantages of optogenetics lie in the spatiotemporal control offered by the system that allows for... expression of ChR2(H134R) bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which...advantage of the extensive supply of transgenic reporter lines that supply a variety of GFP expression ...

...Center (www.taleffectors.com) . Whether you work in a C. elegans lab and have been struggling to mutate a ..., Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic...using engineered TALENs. Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR. Nat Biotechnol...every 35bp. Researchers are still determining the importance of context for each TAL effector within an array...plasmid toolbox. Addgene has rapidly become an important resource for this new technology, and we hope ...

...replicates such as multiple mice in each group and C, technical replicates or repeated measurements will...interactions between the sample and the secondary antibody. C, a label control treats the sample to the full protocol...PubMed Central PMCID: PMC5773976. Waters, Jennifer C. “Accuracy and Precision in Quantitative Fluorescence...binding  Primary antibody specificity is further supported by a pre-immune serum or isotype-specific control...

...80 °C. Titer All titering is performed on lentiviral preparations that have been stored at -80 °C and ...Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or ...expression and/or function. These data are sometimes reported or posted on the material page for the corresponding...recipient's initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...This DNA is then used as a template to amplify a portion of the intended lentiviral insert. During this ...

...come out! References Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C., & Schultz, P. G. (1989). A...describing a technique that allowed for intentional incorporation of an unnatural amino acid in translation (Noren...natural translation machinery; these are often incorporated into the cells via plasmid. Third, a second ...containing the chosen codon that will result in the incorporation of the non-canonical amino acid at the site ...integrated into the cell’s genome, instead of incorporation via plasmid or media. Theoretically, if you ...will produce a protein with a site specifically incorporated ncAA, ready for you do to any number of experiments... They provide resources, knowledge, and most importantly, validated protocols (using readily available...

...Pewe L, Wohlford-Lenane C, Hickey M, Manzel L, Shi L, Netland J, Jia HP, Halabi C, Sigmund CD, Meyerholz...SARS-CoV-2 were developed for broad use and are now important for accelerating research into this virus. One...during the pandemic and in the future, depends on support from the scientific community. Deposits of new ... be deposited into repositories, and financial support to maintain repositories’ capacity for storing,...all ways in which the scientific community can support repositories. Repositories are essential for the...

... a mutation that causes Niemann-Pick disease type C, directly demonstrating the therapeutic potential ...toxicity in long-term experiments.  Figure 4: The C-terminus of Cre (CreC) and the N-terminus of Cre (...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET. Image from Beck...indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo toolkit for... Eisenhoffer lab, is a new tool that increases temporal and spatial precision of gene expression and reduces...

...disorder of glycosylation, and Niemann-Pick disease type C. Although the three DMD studies referenced here represent... 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H, Sanchez-Ortiz...K, Cheng JK, Chew WL, Widrick JJ, Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe...optimized, it’s important to ascertain how long the rescue phenotype will last, and more importantly, if it does...genetic disease in a postnatal mouse model, an important proof of concept for future preclinical and clinical...does not cause obvious germline modification, an important finding given the controversy associated with ...great step forward for CRISPR gene therapy, it’s important to realize that DMD is a simpler case than other...

...also shown that the Q system is also functional in C. elegans. Subsequent work by the Luo and Potter labs...23995928; PMCID: PMC3811519.Seo SO, Schmidt-Dannert C. Development of a synthetic cumate-inducible gene ...; PMCID: PMC5270762. Wendler, F., Park, S., Hill, C. et al. A LexAop > UAS > QUAS trimeric plasmid to ...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...the Q system from a gene cluster in the fungus Neurospora crassa. The QUAS promoter is less leaky than ... transcription from the UAS promoter. This is important because it allows them to be used together for...

...10.1016/j.gendis.2017.04.001 Maetzig T, Galla M, Baum C, Schambach A. Gammaretroviral vectors: biology, technology...2011 Jun;3(6):677-713. DOI: 10.3390/v3060677 Varanda C, Félix MDR, Campos MD, Materatski P. An Overview of...Lenti particles enter the nucleus through nuclear pores and thus do not require breakdown of the nuclear...