...updated in 2023 by Susanna Stroik.  References and Resources References Yuxuan Wu et al. "Correction of a...of the CRISPR-based editing therapies that are currently approved or have made significant progress in ...cells and delivering them to the patient.   Current CRISPR Therapies Cancer and CAR-T therapy The FDA...therapy pipelines (viral), and researchers are currently working on giving this treatment a CRISPR spin...leukemia, lymphoma, and some solid tumors are currently in early-phase clinical trials and have shown ...quite the opposite aim: to destroy. One treatment currently in clinical trials includes Cas3 (the shredder... into many, many pieces. Locus Biosciences is currently running this trial, with more updates expected...

...several different markers, it can also be used to run multiplex assays, allowing the reference sequence...primer  Molecular Biology Reference Molecular Cloning Techniques References Bell AD, Usher CL, McCarroll...each cycle. The raw data is then normalized to a reference sequence present in the same amount in all samples...curve. For CNV assays, it is recommended to use a reference sample with two copies of the gene (Ma, 2014.)...copy numbers, though it has been shown to identify trends correctly (Muhamad, 2011;  Providenti, 2006). Its...probe set for both the gene of interest and the reference sequence. At the end of the run, each droplet ...positive droplets for the gene of interest and reference gene, copy number can be determined through concentration...

... of the plasmids in our repository with known reference sequences, such as full plasmid sequences provided...BLAST, which includes a simple explanation of the different BLAST programs, databases, and elements of the...The blastp and blastx programs are optimized differently and you may want to select one (or both) depending...you want to verify. We will delve into these differences below. Optimizing blastn searches On the Standard... a Sanger sequencing result to a single known reference sequence or to a BLAST sequence database. If you...two or more sequences” checkbox and paste your reference sequence into the Subject Sequence box that appears...BLAST searches. If you do not know the exact reference sequence for your result, choose one of the BLAST...

... in mice Many systemic capsids express differently in different mouse strains, a phenomenon that we are...And these differences occur not just between mice and NHPs; some capsids show both different efficacy ...from AAV9, the vectors have to be administered concurrently to minimize the impact of neutralizing antibodies...determined by the capsid and promoter/enhancer strengths in the cell type of interest as well as the type...from Challis et al. 2022. Image created with BioRender.com. Abbreviations: AAV, adeno-associated virus;...Organism Mouse Strain-Specific Tropism?  Reference AAV-PHP.B* CNS (broad) Mice, rats Yes...Mechanisms of crossing the BBB are indicated in parentheses in the"Mouse Strain-Specific Tropism?" column...

... that differences in gene expression could explain why some ancestry groups were differentially dependent...ancestry-associated genes.   What does the reference genome actually reference?  The CRISPR guides used in the screens...done in this preprint.   Resources and references References Germline variation contributes to false...racial bias, which still impacts and influences current research. Understanding the depth and breadth of... were designed to have a perfect match to the reference genome – even single mismatches within a CRISPR...introduce significant bias into the results. But the reference genome does not represent the diversity of genetic...understand how the use of a primarily European reference genome could affect their analysis. Their work...

...to the plasmid search results: More than 200 different GCaMP containing plasmids are available for distribution...starting a new search or clicking through to a different part of the website, you can now use the pulldown...pulldown menu on the search page to quickly jump to different types of items: Did you notice the “Filter By...you will only be shown filters relevant to your current search. We’ll take a closer look at filters in ...what if you don’t have plasmid ID numbers, or you aren’t sure if any plasmids from a paper have been deposited...James Trimmer Lab published in eLife. Addgene is currently distributing 94 plasmids expressing validated .... The following screenshots show the range of different terms we can use to find materials from this publication...

...discussions on strategies for antibody validation in different applications. Here, I want to highlight the general...proposed antibody validation pillars. Created with BioRender.com.     Orthogonal Strategies This approach...transcriptomic or proteomic data from several different tissues or cell types. If your antibody indeed... signal, so you’ll need a range of expression strengths to be confident in your data. This is a good option...antibodies against the same protein, each targeting a different epitope, and look for correlation between the ...the results. Using antibodies against different epitopes on the same target reduces the likelihood that ...unique antibodies. An additional caveat is that different antibodies may or may not perform well under the...

...19 pandemic,” says Balazs.     References and Resources References Cele S, Gazy I, Jackson L, Hwa S-...vaccination efforts. So, how effective are the current vaccines against the variants? Alejandro Balazs...found that the neutralization effectiveness of current vaccines differed depending on the variants (Garcia-Beltran...plasmids which encode spikes found on all of the different circulating variants. This allowed us to create... of the key drivers in determining whether the current vaccines are effective or not. This protein both... that used live virus (Cele et al., 2021). “We weren't completely shocked [with these results] because...variants. Yet, the B.1.351 variant, which has only 9 different mutations compared to the wild-type, escaped neutralization...

...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies...will help them to understand how different mutations lead to differences in the symptoms experienced by ...with the condition to reveal some “outstanding differences.” They now have some theories on the underlying...Institute and Addgene will make a substantial difference in pushing the work forward. Xeroderma Pigmentosum...lot of those plasmids so that we can try many different things in parallel.” Alternating Hemiplegia of...and ATP1B3 plasmids to explore the functional differences among mutations responsible for the disease known...Childhood (AHC). Patients affected by AHC suffer from recurrent attacks of unilateral paralysis (hemiplegia) that...

... C improved assay reliability. Different primers can have different annealing efficiencies (Wang et ...of samples from different sources should not be compared. If using AAV from different sources in the same...solution, and generally these methods measure different characteristics of the solution. These characteristics...because each sample with be quantified with a different primer pair. Ct values vary based on the amount...and low-concentration samples can also behave differently in PCR, even if both samples fall within the ...reliable AAV titering? For our viral service, we currently titrate AAV vector preparations by qPCR. To ensure...absolute quantification by using the universal AAV Reference Standard Material (AAV SFM), which is an AAV sample...

...intracranial self-stimulation (ICSS) and place preference. The lab I work in (the Kreek lab) focuses on...behaviors to test reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting ...nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement but ...halves that are distinguished visually through different patterns and the animal only receives stimulation...Comparison of c-fos amongst animals that receive different types of stimulation may help to refine and optimize...many known behaviors. However, if you observe no difference in your behavioral output with optogenetic manipulation...manipulation, this can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is...

...experiment uses 2 or 3 different fluorescent colors each measuring a different target. Flow cytometers...analysis. Since different populations of immune cells express unique Clusters of Differentiation cell surface...flow cytometry experiments.       References and resources References: Bonner WA, Hulett HR, Sweet RG, ...each cell as it passes through and scatters in different directions. The degree of scatter is captured ...fluorescent signal that is captured by detectors. The strength of the emitted photons varies from cell to cell...highly versatile and some can accommodate up to 30 different colors. This lends flow cytometry particularly...to complex experiments looking at a variety of different targets. If you are planning on using fluorescent...

...components expressed from different plasmid backbones. They noted that backbone differences such as the order,...assess how different lentiviral plasmid backbone elements impact viral titer and to compare different Cas enzymes...Hurt for his expertise and input.  References and resources References Weber, Ernst, et al. “A Modular ...product in an affordable way, researchers test many different combinations of promoters, terminators, solubility...experimental goals are more general in nature, or you aren’t planning on using K. phaffii, then the MoClo YTK...1: A chart with MoClo kit recommendations for different modal organisms and applications.    Plant...be used to test a variety of Cas enzymes with different PAM requirements and editing abilities, such as...

...Created with biorender.com. Pro tip! If your samples and control underwent different treatment processes...Created with biorender.com. The plot above is for a smeary expression and is quite different from the ...presence or absence, the plots can look quite different. You might have high expression, low expression...shows the population of interest. Created with biorender.com.   In such cases, instead of trying to quantify...yes/no flow cytometry readout. Created with biorender.com.   Using histogram plots to quantify smeary... the mean fluorescence intensity (MFI) of the different populations. MFI is the average brightness from...plot. However, the histogram plots look a bit different than they do for yes-no readouts.      ...

...101 eBook, 1st edition.  References and resources References Swift S, Lorens J, Achacoso P, Nolan GP. ...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...from lentiviruses? There are a few important differences between engineered γ-retroviruses and lentiviruses...lentiviruses. First, these viruses were derived from different genomes (MoMLV and MSCV for γ-retrovirus; HIV ...be useful for the transduction of terminally differentiated (non-dividing) cells. Both lentiviruses and... in gene therapy research to reduce promoter interference and/or drive high expression of a transgene ...

...nucleus, a different fix/perm kit is required (to be precise, the fixing solution is different while the...decrease in signal strength in a flow plot. d = days post treatment. Created using biorender.com. Cell viability...lineage markers, like many of the cluster of differentiation (CD) proteins found on immune cells. However...antibody host species interactions. Created using biorender.com. Another way of indirect labeling is through...primary antibody host species, as it has to be different from the other antibodies in your panel. Otherwise... dendritic cell, CD8+ T cell. Created using biorender.com.   Intracellular labeling Oftentimes, a ...first fixed and then permeabilized, using two different buffer solutions. Fixing causes protein cross-...

...graph of photocurrent vs. time. The resulting histograms is described through maximum current (H), the...cytometer to increase the electric current and thereby the signal strength. So, we need to adjust the voltage...Gates are the parameters the machine uses to differentiate between variations in those factors. To understand...a little bit of theoretical background on the different parameters that your gating strategy will be based...measured: height (H), area (A), and width (W). The different measures are represented as FSC-A/SSC-A, FSC-H...(see Figure 2). H describes the maximum signal strength, while W results from the time that the cell spent...cells, blood cells and more), each coming with a different size and structural complexity. After several ...

...sequence critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced...that amplifies the entire plasmid template. The parent template is removed using a methylation-dependent...A*” (in addition to the “A” site already in the parent vector). Also shown are the “B” restriction sites...with enzyme “A” as compared to the products of parental plasmid digestion (blue). Digestion with enzyme...mutagenesis. In addition, you may want to try different concentrations of template (e.g. 0.1-1.0 ng/μl...methylation-dependent enzyme (DpnI) to eliminate the parent plasmid from the PCR products, the plasmid template...patch up the DNA. The resistance marker from the parental plasmid provides a mean for selecting for transformants...

...conjugated to different fluorophores or a panel of primaries that was raised in different species or are...are complex and antibodies may perform differently in different samples and experimental conditions. Be...for IHC, not all IHC experiments are the same. Differences in sample processing, fixation, antigen-retrieval...make sure that you can easily distinguish each different type of antibody and therefore each target protein...unique isotype or species, then you can choose different fluorophore-conjugated secondary antibodies specific...product. Note that many antibodies are sold under different names through multiple vendors, so look at epitope...to choose between the same antibody sold under different names.  If all else is equal, feel free to make...

...expression of kynurenine aminotransferases, to enhance the conversion of kynurenine into kynurenic acid. That...skeletal muscles led to another clue: differences in the kynurenine pathway of tryptophan degradation. It...That’s key because, unlike kynurenine, kynurenic acid can’t cross the blood-brain barrier. By lowering...plasmids from Addgene. References: Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates...lowering circulating levels of kynurenine in the body, the brain is protected from stress-induced biochemical...