We narrowed to 965 results for: REP

... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...considered hypotriploid: most chromosomes are represented by two to three copies, but several have four...your research question, HEK-293 is known for reproducible results. It’s also widely available, which can...biology, ion channels, or involve human protein preparations with PTMs. Why the HEK not? References and ...

...plasmid. Inverted terminal repeat (ITR) sequence Inverted terminal repeats (ITRs) are what makes an AAV...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...(2019) A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods...

... posts of 2015 1. CRISPR 101: Homology Directed Repair Addgenie Chari Cortez explains some of the basic...basic science behind homology directed repair (HDR) and how it’s used in CRISPR genome editing.   2. Plasmids...While the vast majority of CRISPR plasmids in our repository use Cas9 to cut DNA, Addgene Science Communications...explain some of the molecular gymnastics behind DNA repair by non-homologous end joining (NHEJ). 7. Career...Resources Learn More about Viral Vectors Browse the Repository  ...

... about sexual and reproductive health, as the stigma around sexual and reproductive health issues creates...to all the optional sexual health sessions and reported the intel back to my shyer classmates. For my ...evaluate the quality of the study those abstracts represented. The point of this study was to see if their ...research focuses on misinformation about sexual and reproductive health.    References and Resources References...

...Gerald Schwank, first author of the cystic fibrosis report. “This is a big advantage over induced pluripotent...also need to be perfected until the efficiency of repair approaches 100 percent. “We need to be sure it'...it's accurate enough and efficient enough for repair,” Doudna said. “And it can't be too hard to apply....need to get a handle on how cleavage sites are repaired.” Those insights will be critical for scientists...December 2013). 8. Gerald Schwank et al. "Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell ...

...You’ve prepped your DNA and you’re ready to get started on the next step of your experiment. But in many...give you an idea of the concentration of your DNA prep and whether there are any other contaminants. Nucleic...your sample against this curve to quantify your DNA prep. Though this method takes more time to set up in...and is not used as much for the standard plasmid prep. Diphenylamine method Another absorbance based method...Learn about the different grades of plasmid DNA preps Resources on Addgene.org Visit the Molecular Biology...

...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...Detailed documentation, which facilitates the reproduction of the structure in a similar way to the biological...assemblies open-source. The sharing of digital representations of the 3D models, which in many cases can be...with the goal of creating more transparent and reproducible science in the long run (Hohlbein, et. al., ...

...enough to get it into cells and let them do the repair work. The key is T4 DNA polymerase, which has 3...forming a circular product with four nicks easily repairable by the bacteria after transformation. LIC is ...transformed into bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   ...intermediate is transformed into bacteria, which repair the plasmid. Created with BioRender.com. How...PCR-generated inserts. By harnessing the power of DNA repair, you can assemble multiple fragments without the...

...plays an integral role in regulating genome replication, repairing mismatched basepairs or small indels that...occur during DNA synthesis, and protomoting or repressing protein expression. Methylases involved with ... (as these would also be deficient in mis-match repair functions of Dam), so these strains should not ...Learn to Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources ...

...knockout screening). As the Zhang lab's Nature paper reports, “Here we have shown that the SAM system is robust...investigation of the cellular consequences of repressing or inducing individual transcripts. The activator...site in 15,977 human genes. The transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each ...be turned on and off, to reversibly activate or repress genes of interest. They also “showed that different... Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson...

...proteins known as ankyrin repeat proteins. DARPins typically consist of 3-4 repeats, have high specificity...instead of generated in vivo, they have higher reproducibility and consistency in the final product. Because...refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically engineered...combinatorial libraries, and high specificity has been reported for a number of targets. Due to their size and...

...Primary antibodies are represented in blue; secondary antibodies are represented in purple; signaling molecules... molecules are represented in light green; and proteins are represented in orange. Created with BioRender.com...architecture of the indirect approach is as follows: Prepare the sample. Incubate the sample with the primary...

...catalytically dead Cas9 fused to a transcriptional repressor to your gene of interest. It will inhibit transcription...more on this below) will both achieve temporary repression of protein expression. Addgene has a dCas9 KRAB...CRISPRa blog! Fig. 1 CRISPR activator and repressor mechanisms Want to transiently induce expression...suited for the job. Whether you want to express or repress, good luck on your protein expression journey! ...Plasmid 101: Inducible Promoters Plasmid 101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose...

...including a handful of plasmids that have been in the repository for many years, has now grown into a collection...neutralization assays, part of a study that examined the repertoire of antibodies from 19 COVID-19 convalescent individuals...tools based on Cas12a and Cas13  which could be repurposed for SARS-CoV-2 RNA detection. Early in the pandemic...above, some of these plasmids have been in the repository for many years while many were developed during...Harrison SC, Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike ...

...that meant rereading many of the original papers reporting various prime editing tools. These papers are ...quick refresher: prime editing is a “search and replace” genome editing technique that relies on a fusion...or longer) tracts of edited bases. DNA mismatch repair (MMR) is less efficient at identifying these “bubbles... use the edited strand as a template during DNA repair. Test multiple nick sites, starting with sites ..., Raguram, A., & Liu, D. R. (2019). Search-and-replace genome editing without double-strand breaks or ...

...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although... inherited by daughter cells during cellular reproduction, similar to the host’s chromosome, or they can...For example, helper plasmids were developed to replace helper viruses in AAV production systems. We’re...the many plasmid tools available in the Addgene repository. To learn more about some other common plasmid...plasmid features, such as origins of replication and promoters, check out our other Plasmids 101 posts, and...

...Associate Professor at Boston College. National reports stress the importance of providing authentic research...overexpression vector. This plasmid contains origins of replication and selectable markers that allow students to...functional MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes...complementation should only occur when galactose replaces glucose as the carbon source. Student results .... cerevisiae met deficiencies (manuscript in preparation), indicating that Met protein functions have ...

... on the composition of the viral vector and the repeat sequences present in the LTR, the vector may undergo...undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying the... and the type of virus, there have been varied reports of titer losses from 5% to 50% per freeze thaw ...exchanging the medium after transfecting cells, replace the medium with half the volume typically used ...virus particle to a target cell by reducing the repulsive electrostatic forces between these two negatively-charged...

...are difficult to modify through homology-directed repair. Both Cpf1 and its guide RNAs are smaller than ...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions... may improve the frequency of homology-directed repair (HDR) over non-homologous endjoining (NHEJ). Cas9...chance” mechanism. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Many ...

...modification with Homology Directed Repair (HDR), CRISPR activation or repression, transgenic mouse line creation... which means they are non-infectious and non-replicative. Nanoblades are produced by transfecting the ...compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades CRISPR-based HDR allows for...transduction efficiency by neutralizing the charge repulsion between virions and the cell surface, and also...