We narrowed to 853 results for: Eras

...year and how these plasmids have been used to accelerate COVID-19 research in other labs. Spike protein... protein has had many opportunities to evolve. Several variants are now circulating and plasmids containing...from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding different spike...used these pseudoviruses to determine how effective sera from vaccinated individuals are at neutralizing ... the reverse transcriptase, protease, and Bst polymerase constructs from their assay to Addgene.  Drew...expressing MMLV reverse transcriptase and Taq polymerase that can be used in a one-step RT-qPCR reaction...SARS-CoV-2 researchers to investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids...

...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase increases levels...transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and epigenetics ...initiates cytosine demethylation of DNA. However, several proteins in the DNA oxidation and repair pathways...delivery systems. Transcriptional repression DNA Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited...repression. Created with BioRender.com. DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1...Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase from the prokaryote Mollicutes spiroplasma...

...human gene therapy. Molecular Therapy: The Journal of the American Society of Gene Therapy, 14(3), 316...the viral particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor...strains of one species, so be sure to check the literature and validate your choice of AAV vector for your...AAV1 was the first viral vector approved for gene therapy (Issa et al., 2023). It may be a good choice for...central nervous system, including neurons, AAV5 is generally the best choice for targeting astroglia. It has...skeletal muscle, although muscle inflammation and degeneration has been reported (Muraine et al., 2020). AAV8...depends on many variables, so be sure to check the literature for previous work in your experimental system...

...There are several facilities nationwide where you can obtain excellent quality, high titer rAAV (including...including Addgene!), but you can also generate rAAVs in your own lab with standard molecular biology tools...our ddPCR for AAV quantitation blog post.  We generally do not recommend using ELISA-based titering assays...inverted terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org/10.1089... of gene expression in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https://doi.org/10.1186...can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038/sj.gt...Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. https://doi.org/10.1089...

...providing many of the details not generally captured in the literature, scientists can use the Data Hub...commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists... process is often not detailed in the primary literature, and as a result can leave scientists under-informed...time and effort to test and optimize our vector - overall, I spent two years testing and reformulating our...dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https://doi.org/10.1038/s41434... Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089/hum.2009.169...

...the functions of different genetic elements to generate non-natural tools such as novel biological circuits...and more. By combining these components, one can generate constructs to visualize cell cycle dynamics, titrate...applications. In one example, modular constructs generated with MXS-chaining were used to label subcellular..., MXS Chaining gave researchers the ability to generate the four initial unique constructs and later combine...ligation, the original pattern of the MCS is regenerated at the flanks of each ligation product, which...that the restriction sites of the MCS are only regenerated at the 5’ and 3’ termini of the ligation product...added to the 5’ or 3’ terminus using the newly generated MCS. MXS-chaining also has the advantage of being...

...FLEx cassette using next-generation sequencing (NGS) (Guerin, K. 2020). In general, we observe that 0.1 -... in the Central and Peripheral Nervous System and Relevance to Human Gene Therapy. Front Mol Neurosci ...FLEx Vectors Important Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting...Vectors). While this strategy greatly reduced the overall level of off-target transgene expression, low levels...greater off-target expression just due to more overall viral particles getting into those cells. Therefore...transduces your cells of interest. We recommend testing several different serotypes to determine which one is the...course experiment to check expression levels over several weeks allows the researcher to determine when the...

...begin to use CRISPR as a therapy. Low frequency off-target effects may generate irreproducible data in ... CRISPR experiments, many are turning to next-generation sequencing options over older assays. Biased... search criteria are not exhaustive. Mismatch tolerance settings are often limited to off-target sites...sites of <4 bp. The off-target list is also generally weighted by the position of the mismatch along the ...CRISPR editing in the model cell line or organism. Several options exist for unbiased off-target detection...labeling, enrichment on streptavidin and next-generation sequencing (BLESS) (Crosetto et al. 2013). GUIDE-seq...consistent set of standards as the field seeks to generate reproducible, quality data on the role of genetic...

...plasmids. Growth conditions The amount of aeration, temperature, culture volume, antibiotic, and medium ...survive within cells. The replicons of plasmids are generally different from the those used to replicate the... to make additional copies. ori sequences are generally high in As and Ts. Why, you ask? Well, A-T base...pairs can be separated more readily at lower temperatures and allows the replication machinery room to...few copies depending on how they are regulated. Generally, control of replication is referred to as "relaxed...exclusively in E. coli? Gram negative bacteria in general? Both Gram negatives and Gram positives? Will you...compatibility with one or more other plasmids. Generally speaking, plasmids with the same ori are incompatible...

... We’ve previously discussed some of the general considerations for performing behavioral tests, but now... modulating. Viral vectors, in contrast, are considerably more versatile and specific in how they manipulate...field box or a circular corridor apparatus using a camera or tracking software. Locomotion is a highly complex... of individual neurons and in terms of their interactions with one another. In addition to the neuronal...controlling muscle activity and coordinating movement, several additional brain areas that reflect motivational...sensitization and might open new ways for a novel therapeutic approach against drug addiction. Anxiety tests...rodents can be used to study anxiety levels in general, or anxiety associated with psychiatric diseases...