We narrowed to 735 results for: RAN-1

... inhibitory synapses in the brain.  Figure 1:Schematic diagram of different FingR constructs and... efficient double stranded templates, rather than the more efficient single-stranded oligodeoxynucleotide...generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent proteins... them to localize to the synapse and minimize extraneous fluorescence from other regions of the cell. ...synaptic maturation pattern of hippocampal adult-born granule cells over time that were not previously possible... al., 2020). Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations...lentiviral CRISPR library that targets human glycosyltransferase genes and other proteins related to cellular...

... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...this surface”….where the complex is docked on the membrane of the endoplasmic reticulum. And graduate students...reported cleavage site on the complementary DNA strand. And this is why we love working with researchers...”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting physical...protein must have in order to scan along the double-stranded DNA, recognize a PAM sequence, allow the complementary...pairing between the guide RNA and the target DNA strand, and then activate two different endonuclease active...structure captures the displaced non-target DNA strand and shows the HNH nuclease domain swung around ...

...article, they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit from...spatially and temporally target single cells. Now, Francois St-Pierre’s lab developed a new form of FACS-based...-based Dual-fluorescent DSB Repair) is a double-strand break repair assay for mammalian cells. It's based...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced...

... little signal detected along the neurite (Figure 1.) Moreover, the group was able to demonstrate that...expression in E. coli. This operon is under the transcriptional control of a strong constitutive promoter (... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover... protein kinase A (PKA) activity at the plasma membrane in living cells using a construct in which TagRFP-T...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy...

...technologies (Bradbury & Plückthun, 2015a).   Figure 1: (Top) Polyclonal antibodies are produced in animals...chain and light chain for the antibody and when translated into protein will assemble into a fully functional...synthesize the genes that encode those amino acids (Tran et al., 2016). Each antibody would then be tested...P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018) When monoclonal antibodies...:633–644 . https://doi.org/10.1083/jcb.201409074 Tran NH, Rahman MZ, He L, Xin L, Shan B, Li M (2016) ...

...your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified...would at least be guaranteed a successful result? While there are never any guarantees, some cloning techniques...Resources” section at the end of the post! Speed: Your grant deadline is approaching or your advisor is demanding...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...

...most comprehensive for searching. Timesaving Tip #1: If you know the species that your sequencing result...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

...SpCas9 base editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated...Lower on-target activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very ...scientists to quickly and easily generate zebrafish transgenic lines to study neurobiology and neuorodgenerative...pluripotent stem cells) through the expression of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the... method for generating iPSCs to express the transcription factors from a polycistronic cassette. However...comparable experimental results. Using PiggyBac transposons, the Woltjen Lab compared different variants ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at ...

...cereblon, leading to rapid target degradation (Figure 1). Plasmids: We generated a series of plasmids that...example, IZKF1 and IZKF3 are cell-type specific transcription factors with important roles in Multiple Myeloma... (Nabet et al., Erb et al., Huang et al.), transcriptional and epigenomic profiling studies (Nabet et ...Featured molecules: dTAG-7 and dTAG-13 Erb et al. Transcription control by the ENL YEATS domain in acute leukemia...

...which you’ve already fluorescently labeled. Figure 1: Using Split-BioID to Identify Protein Protein Interactions...Interactions Difficult to detect weak and/or transient PPIs. Many PPI identification techniques rely ...often prevents the identification of weak and/or transient interacting proteins. No spatiotemporal control...BioID, is also useful for identifying weak and/or transient protein-protein interactions since the biotin ...

...Set a goal for paper-reading, even starting out at 1 paper/week will be useful. Check out the hashtag #365papers...your area. When you are in ‘writing-mode’ for a grant or a scientific manuscript, the reading will likely...

... to the use of multiple antibodies. Figure 1: Overview of using Sandwich ELISA to quantitatively...concentration of any experimental wells in that range of optical densities. If your standard curve does...

...significant progress in clinical trials.     Figure 1: Types of delivery and editing strategies for CRISPR...CRISPR targets — Sickle Cell Disease (SCD) and Transfusion-dependent beta thalassemia (TBT), both of which...clinical trial for SCD in 2022 as well. Hereditary transthyretin amyloidosis (hATTR) hATTR is caused by a variety...

...editing machinery such as CRISPR-Cas. Figure 1: Schematic of targeted gene knock-out (left) and targeted...AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed cells...(right) are transduced to target cells. Within target cells, Cas9 machinery is transiently expressed and... viruses. Adenoviruses are non-enveloped double-strand DNA viruses with a genome of around 36 kb in size...left), a fragment is deleted from two homologous strands of DNA through error-prone non-homologous end joining...other viral vectors, AdVs can provide a large transgene packaging capacity. For instance, with a packaging...disruption frequencies in the various cell types ranged from 18% to 65% (Maggio et al., 2014). An alternative...

...Figure 1: A schematic illustration of CRISPR/Cas9-based approaches to study ncRNAs. (A) Transcription start...signal downstream of the transcription start site will terminate transcription of most of the ncRNA (Figure... coming, right? Non-coding RNAs (ncRNAs) are transcribed but do not encode proteins. They play important...Using a single guide RNA (sgRNA) to create a double-strand break works well because very small deletions and...sequence can completely disrupt expression of the transcribed ncRNA (Jiang et al. 2014).  Simple knock-out ...lncRNA regulatory elements that are crucial for transcription and downstream function. Rather than attempting...the lncRNA locus, scientists can mutate the transcription start site, splice sites, exons, and/or promoters...

...how the brain operates. References 1. Callaway EM. Transneuronal Circuit Tracing with Neurotropic Viruses...generally are not transmitted to adjacent neurons via synapses (i.e., transmitted transsynaptically). This is ...retrograde transmission (Etessami et al., 2000, Wickersham et al., 2007). Thus, RABV transsynaptic spread ...replicate. Modified Rabies Virus: Converting a transsynaptic neuronal tracer into a monosynaptic (first order...subsequent cycles of viral replication and retrograde transmission occur, spreading the virus through higher and...higher order neural connections. While this transsynaptic spread is useful for neuronal tracing, it has...-dependent TVA to Cre-expressing neurons in a transgenic mouse line. By doing this, RABV expressing the...

...genome engineering and microbiology.     References 1. Chen, Sean, et al. "Highly efficient mouse genome...:     Cells must be amenable to transfection or viral transduction     Appropriate promoters must be...expression     The requirement for Cas9 transcription and translation delays editing What are Cas9-gRNA ... studies in cells that are difficult to transfect or transduce. While using Cas9-gRNA RNPs may require...site of interest. This is commonly performed by transfecting cells with a plasmid, such as PX459, which encodes... often be used in cells that are difficult to transfect, such as primary cells. Using RNPs can also alleviate... are detectable at high levels shortly after transfection, and are quickly cleared from the cell via protein...

... NGT PAM GAA PAM GAT PAM xCas9 3.7 41 (1) 32 (1.6) 13 (2.1) 38 (4.5) 7.2 (5.2) ...xCas9 for transcriptional regulation and base editing Hu et al. also engineered transcriptional activators...activating transcription. Sites containing NG, GAA, or GAT PAMs averaged 56-91% transcriptional activation... to create xCas9, an enzyme recognizing a broad range of PAMs like NG, GAA, and GAT, but also displaying...subunit ω. This construct is designed to activate transcription from the accessory plasmid (AP) encoding phage...could recognize a library PAM, it would induce transcription of gene III, permitting phage propagation and...spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing NG, NNG, GAT, and CAA PAMs, while xCas9...

...target DNA quickly and accurately.  Figure 1: Steps in PCR reaction   This process of denaturation...separation of the two DNA strands.  Annealing- The temperature is then dropped to a range in which the forward...the primers can anneal to the single stranded DNA template strands. DNA polymerase is also stable at this...template DNA to amplify from and unique single stranded DNA primers that bind upstream (5’) and downstream... process as DNA polymerases require an existing strand of DNA to add nucleotides to.  Using these reagents...reaction:  Denaturation- To amplify DNA, the two strands of the template DNA first have to be separated....used to quantitate RNA levels using reverse transcription-qPCR (RT-qPCR). The first step of this process...

...in restraint. The very first subheading in Chapter 1 asks “What is science?” That’s a very big question... that may have warranted their own chapters, and certainly chapters that could warrant their own books..., and he wisely makes no such attempt. He is transparent about his own biases--many of which will be shared...non-scientists, Zimmer’s observations will likely range from mildly surprising to genuinely shocking. Mainstream...he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in the identification...within this section. The GFP story has its own Franklin in Doug Prasher, who originally isolated the GFP...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...