We narrowed to 432 results for: sequence

...targeting power of CRISPR combined with the large sequence capacity of transposons open whole new worlds ...cofactors. The transposon proteins integrate a payload sequence into genomic DNA downstream from the Cas target... double-stranded breaks and can deliver large sequences and even entire genes. Figure 1: Mechanism... transposon proteins (Tns) to integrate large sequences into genomic DNA downstream of gRNA-specified ...site-specific recombinase to achieve insertion of long sequences. In their experiments, PASSIGE had higher efficiency...

...viruses. Evolving to survive Although the CRISPR sequences were initially discovered in E. coli in 1987 (... (1), the concept that these clustered repeat sequences operated as a safeguard against bacteriophages... 30bp in length and paste them into the CRISPR sequence. Separate Cas proteins then express and process...using slightly different Cas proteins and PAM sequences. A total of eight evolutionarily conserved CRISPR... generating a single point mutation in the PAM sequence, preventing the Cas nucleases from re-identifying...

...codons that alter the nucleotide sequence but not the amino acid sequence of the translated protein.  These...however, one of its biggest limitations is that sequence modifications can only be made at restriction ...and deletion of selectable and non-selectable sequences, point mutations or other small base pair changes...recombination of the substrate with the target DNA sequence. This means cloning occurs in vivo, as compared...made by PCR using primers that amplify the DNA sequence of interest and flank it with 50 base pairs of...nucleotides long (20 nucleotides that anneal to the DNA sequence of interest and 50 nucleotides of homology to ...desired alteration(s) located in the center of the sequence. Since lambda red has a higher recombination frequency...

...mismatch between the original sequence on one strand and the edited sequence on the other strand. To guide...transcriptase with a sequence optimized for human codons; additional nuclear localization sequences; and two mutations.... After nicking by Cas9, the homologous pegRNA sequence hybridizes to the target site, and the reverse...Advantages of prime editing Less constrained by PAM sequence location The prime editor extends the reach of...the PE2 enzyme. PEmax: Optimizing prime editor sequence to improve expression and activity In addition...  ✓ SV40 and c-Myc nuclear localization sequences Improve translocation to the nucleus ~ ~...prime editing with a recombinase to install large sequences (up to 10 kb) into the genome (Pandey et al., ...

...the gene of interest. This gives you loads of sequences to select from - in many cases several exons’ ...sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the genome.  ...upstream of a Protospacer Adjacent Motif (PAM), the sequence NGG.  If these conditions apply to your experimental...feasible for your experiment, ensure your sgRNA sequence has little homology to other regions of the genome...polymerizes DNA onto the target strand based on the sequence of the pegRNA. Image from David Liu.   Activation... sgRNA vectors and design your sgRNA targeting sequence using one of the many freely available sgRNA design... cells, it is time to confirm that your target sequence has been modified!  The exact protocol may vary...

...simultaneously  With increasing sequencing power, scientists are able to quickly sequence an individual’s genome... develop a method that measures the effects of sequence variants on abundance simultaneously called VAMP-seq..., for Variant Abundance by Massively Parallel sequencing. In VAMP-seq, the abundance of a protein variant...Genomic DNA from each bin are then extracted and sequenced. Chiasson is currently using VAMP-seq to study...

...different synthetic peptides corresponding to linear sequences within the middle of the human c-Myc protein or...chicken homologs. By comparing the homologous sequences in these species, they found that the N-terminal... as a recombinant antibody. Researchers first sequenced the variable domains of Myc1-9E10 in the late ...affinity for the epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). ...Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al.’s results...variable epitope recognition dependent on neighboring sequence context. Sci Signal 13. https://doi.org/10.1126...

...orthologs, Cpf1 efficiency varies widely with gRNA sequence. Both groups used multiple methods to examine ...single and double mismatches throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except... they were present in the 3’ end of the target sequence (bases 19-23). Cpf1 is also sensitive to single...23. Accordingly, the 3’ end of the gRNA target sequence does not have an essential function in Cpf1-mediated...4-6 base deletions at the 3’ end of the target sequence. Cpf1’s strength may lie in its low off-target...post-HDR, repair templates should remove the PAM sequence. Many research and clinical applications require...

...discrepancies between what we sequenced in house and the submitted sequences were a concern to the plasmid... currently working on the QC/QA team as a DNA Sequencing Intern. The application process When I first...approachable.     Fig. 1: Examples of DNA sequencing QC work    In addition to my responsibilities...

...majority of information on each cancer clinical sequencing report includes rare variants that haven’t been...corresponding to cancer variants found in genome sequencing studies (as well as reference cDNA clones mapping...scientific community through Addgene. These include 634 sequence confirmed mutant cDNA clones corresponding to ... worked with many collaborators to aggregate a sequence-confirmed series of clones corresponding to known...when rare variants are discovered in clinical sequencing studies. At the Broad, my colleague Cory Johannessen...