We narrowed to 803 results for: Cre

...Cancer cells find a way to grow haywire, access and create more vasculature to feed themselves, use the blood...the tumor microenvironment - a vital atmosphere created by the cancer cells to shield themselves from attack...cells of epithelial origin. Suppressing function, creating tolerance, and evading recognition are a few methods...cells into regulatory cells to suppress function. Creating Tolerance T-regs in the tumor microenvironment...HIF-1), which promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates...signals released from the microenvironment help create tolerance beyond direct proximity to the tumor;...cells (DCs) and macrophages (M1 and M2) that help create welcoming environments for tumor cells spreading...

... KJ’s successful base editing intervention was created. Only about half of babies with KJ’s disorder survive...survive long enough to receive a liver transplant — creating intense time pressure for the research team. Ultimately...base editing advances that led to KJ’s treatment. Created with BioRender.com. Invention of base editing...DNA-targeting technology with a fused cytidine deaminase to create C→T base edits at a target locus. A year later,... (Gaudelli et al., 2017). These editors did not create double-stranded breaks in the DNA backbone, making... be targeted (Walton et al., 2020). The initial screen for base editors that could target KJ’s mutation...PAMs available, a large number of gRNAs could be screened for the most efficient editing. In these initial...

...mutations increased the accuracy of editing by up to 10-fold per allele. This is a 100-fold increase in editing...incorporate homo- or heterozygous mutations. 1. Increasing HDR accuracy with CRISPR/Cas-blocking mutations...find the 0.2-0.5% correctly edited alleles, when screening reasonable amounts of single cell clones. Furthermore...tedious business of iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000... together with the intended sequence change and screen a few hundred clones to find the right one Step... few hundred. From these two steps, you’ll be screening a total of a few hundred clones - Still better...efficiency of mutation incorporation falls with increasing distance from the cut site. If the distance is...

...recombination, increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability...that make major improvements such as those that increase plasmid yield and/or DNA quality. Table 1 below...sensitive. dnaJ Mutation in a chaperonin gene Increases the stability of certain expressed proteins ...methylate DNA lac Lac operon mutations Blue/white screening of clones   lacIq lac repressor overproduced...derivative. General cloning and storage, blue/white screening, leucine auxotroph. F- endA1 recA1 galE15 galK16...cloning and storage of common plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR...General cloning and plasmid maintenance, blue/white screening, partly restriction-deficient; good strain for...

...encoding Gag and Pol, increasing the number of recombination events necessary to create a replication-competent...Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...shows how the lentiviral genome was condensed to create the 2nd-generation lentiviral system (Figure 2)...safer than the original system, the possibility of creating a replication-competent virus via recombination...possibility and enhance biosafety, Dull et al. created the 3rd generation system (Figure 3), which differs...Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the lab - the Trono lab’s lentiviral...both for targeted modification and genome-wide screening.   Lentiviral vectors in the clinic Since lentiviral...

...CBEs). But unlike CBEs, CGBEs then excise the U to create an abasic (empty) DNA site using either fused or...nucleobase component of the nucleoside inosine. Created with BioRender.com. Inosine excision leads ... able to edit A → Y (Y = T or C) in 67.17% of screening cells (Tong, Wang, et al., 2023). Based on this... for A → X base editor (X = any nucleotide). To create a more specific A → C base editor (ACBE), they ...Black arrows indicate natural cellular processes. Created with BioRender.com. The first deaminase-free...either to directly accomplish the base edit or to create the substrate for a base excision repair pathway...nCas9. First, MPG removes the G base from DNA to create an abasic site. Like in most other base editor ...

...Marker B. Created with biorender.com.   The goal of flow cytometry is to use increasingly more stringent...populations that are B+ and one population that is B-. Created with biorender.com.   If it’s hard to conceptualize...imagining (or drawing) a quadrant over the dot plot. Created with biorender.com.   Of course, this is quite...al., 2017 to identify mDCs and pDCs. Used under Creative Commons license.    In the third plot, anti-CD14...visualize the different populations. Used under Creative Commons license.    Pro tip! Subtypes of immune...the percentage of cells in each gate. Used under Creative Commons license.    Contour plots  FACS data...data from Jhunjhunwala et al., 2015. Used under Creative Commons license.  Histograms  For analysis...

...in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes. For... storm. While Cas9 has been most widely used to create specific edits in DNA, there has also been significant...inducible upon environmental triggers. This is done by creating a split dCas9 protein that can reassemble when...the best uses for Cas9 activators is in genetic screening. gRNAs targeting every gene in the human genome...interactions. Additionally, large scale CRISPR-based screens can be used to gain insight into protein structure...focused advancing scientific discovery through the creation of innovative technologies. To date his group ...of cancer therapeutic targets using CRISPR–Cas9 screens. Nature 568:511–516 . https://doi.org/10.1038/s41586...

...nuclease domains, researchers created the CRISPR nickase. Nickases create a single-strand rather than a...generated by the nickases; D10A creates 5’ overhangs and H840A creates 3’ overhangs in a PAM-out design...Conversely, the H840A mutation in the HNH domain creates a non-target strand-cleaving nickase. Instead of...strands bluntly with WT Cas9 and one gRNA, you can create a staggered cut using a Cas9 nickase and two gRNAs...other? The gRNAs must target different strands to create a DSB, but this can be accomplished with either...an individual gRNA with WT Cas9, repair levels decrease rapidly 10 bp from the cut site. So if you can... you can’t obtain high HDR efficiency. Nickases create a staggered cut - could this system mediate repair...

...autophagosomes with lysosomes leads to an increase in yellow puncta but a decrease in red puncta. Find the mRFP-...RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence suggests an increase in autophagic...flux, while an increase in GFP fluorescence relative to RFP fluorescence suggests a decrease in autophagic...yeast or the lysosome in mammalian cells. This decrease in pH quenches SEP but not DsRed, leading to emission...Therefore, the formation of autophagosomes leads to an increase of yellow puncta (GFF positive/RFP positive), ...fusion of an autophagosome with a lysosome. An increase in autophagy leads to more yellow and red puncta...