We narrowed to 430 results for: sequence
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TypeBlog PostPublishedMarch 3, 2020, 2:15 p.m....reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme. Each half of the base... cell. Intein splitting removes any exogenous sequences and regenerates a native peptide bond at the split...markers, transformation acceptor vectors, and sequences for genomic integration by homology directed repair...
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Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering
TypeBlog PostPublishedMay 3, 2018, 1:48 p.m.... a protospacer flanking sequence, so you can target virtually any RNA sequence! In bacteria, Cas13d-mediated...The Cas13d proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs... -
Viral Vectors 101: Viral Applications
TypeBlog PostPublishedApril 13, 2023, 1:15 p.m....: shRNAs are short RNA sequences which target complementary mRNA sequence for degradation. Viral introduction...viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA targeting... -
Plasmids 101: Yeast Vectors
TypeBlog PostPublishedFeb. 25, 2014, 7:11 p.m....These vectors contain an Autonomously Replicating Sequence (ARS) derived from the yeast chromosome. As the...part of an ARS along with part of a centromere sequence (CEN). These vectors replicate as though they ...DNA (apparently independent of a known specific sequence) dictate the replication of these vectors. S. ... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m....HHCS-associated sequences had lower IREB2 binding enrichment compared to wild-type sequences and that loss...however, knock-in of BoxB sites into endogenous DNA sequences could allow one to study RNAs expressed from their... -
Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9
TypeBlog PostPublishedDec. 16, 2015, 3:30 p.m....lowering Cas9 expression, and truncating gRNA sequences used for targeting; however, these options can...the gRNA (see figure below) Because off-target sequences have less complementarity to the gRNA, they have...Lab similarly thought that if they could weaken sequence independent interactions between Cas9 and DNA,... -
Experimenting in an Open Source lab: from CRISPR to Cats
TypeBlog PostPublishedJuly 23, 2015, 2:30 p.m.... available, and the collection of samples and sequencing is being organized and funded by someone else...the makerspace comfortable with next-generation sequencing data and the bioinformatics pipeline. It’s possible... the Illumina GAIIx we recently acquired for sequencing again - it’s too inefficient and the reagents... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....N-terminal consensus sequence (MG-) and prenylation requires a C-terminal consensus sequence (-CaaX). Both modifications...modifications are disrupted when GFP is fused to these sequences. When N- and C-terminal fusions are not tolerated...alternative is to insert the GFP into the coding sequence of the protein of interest. One of the reasons... reaction requires molecular oxygen and as a consequence GFP remains non-fluorescent under anaerobic conditions... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m....also the first eukaryotic genome to be entirely sequenced and is very amenable to genetic manipulation. ...organisms, the C.elegans genome is also completely sequenced and over 60% of human genes have an ortholog in...Kerstin, et al. "The zebrafish reference genome sequence and its relationship to the human genome." Nature496.7446...extrachromosomal maintenance and integration of transforming sequences." The EMBO journal 10.12 (1991): 3959-3970. PubMed... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog PostPublishedApril 9, 2019, 1:13 p.m.... Since researchers can generate almost any DNA sequence in the way that they like, the design and engineering...mNeonGreen, mScarlet(-I), and mCherry. Keep the linker sequence short In the early days of GFP applications, many...site and therefore consist of random amino acid sequences or (ii) designed to form an inert, unstructured...and protein of interest (POI). To indicate the sequence, people often use terminology like “C-terminal...interest usually outweigh potential negative consequences of the tag. Many thanks to our guest blogger... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog PostPublishedApril 24, 2015, 2:39 p.m....segments, each with two fluorescent protein coding sequences in opposite orientations. Cre-mediated excision...the system, the fluorescent protein whose coding sequence is closest to the promoter will be expressed. ...used are fluorescently distinct, their protein sequences are highly homologous, preventing design of antibodies...three with low levels of both fluorescence and sequence overlap (coral mOrange2, jellyfish EGFP, and sea... -
Plasmids 101: Common Lab E. coli Strains
TypeBlog PostPublishedNov. 7, 2014, 2:56 p.m....MG1655 Wild type K-12 strain; first published sequence of an E. coli K-12 strain. F- λ- ilvG- rfb-50 ...and retroviral vectors or cloning or repeated sequences with the potential to recombine. F' proA+B+ lacIq...and retroviral vectors or cloning or repeated sequences with the potential to recombine. F- endA1 glnV44...and retroviral vectors or cloning or repeated sequences with the potential to recombine. This strain is...E.coli strains Gene(s) Description Functional Consequence dam DNA adenine methylase mutation (GATC) Preparing... -
Your Top Requested Plasmid in 2016!
TypeBlog PostPublishedJan. 11, 2017, 8:47 p.m....strange breaks at target sites 5' of the 5'NGG3' PAM sequence as directed by a gRNA. 2A-Puro In this plasmid...into which you can clone your 20 nucleotide guide sequence of choice (for generation of a gRNA that will ... -
Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)
TypeBlog PostPublishedJuly 26, 2018, 12:46 p.m....phosphorylatable residues and the bNLS and NES sequences, which have previously been characterized. See...more details on the design of the bNLS and NES sequences. Disadvantages of KTRs KTRs also have some disadvantages... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m....that recognize, cut, and alter specific target sequences in DNA resulting in a variety of outcomes including... requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where B is G, T, or C). The authors...scientists seeking a wider range of targetable genomic sequences could benefit from CRISPR-Casɸ. Pausch et al.... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog PostPublishedMarch 12, 2020, 1:15 p.m....strategies: Strategy #1 (ProA): Create promoters with sequences upstream of the start codon of retinal cell-type...Create promoters using phylogenetically conserved sequences that are upstream of the transcription initiation... factor binding sites interspaced with random sequences. Strategy #4 (ProD): Create promoters with the... -
Pathways Over Time Plasmids Engage Students in Functional Genomics Research
TypeBlog PostPublishedMay 22, 2018, 1:38 p.m....In the O'Connor lab, we recognized that genome sequencing projects were generating huge amounts of data...available. Then, staff or students clone similar sequences from a less-well studied organism into overexpression...experiments We used TOPO cloning to insert the coding sequences for ten different MET/Met genes (Figure 1) from... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog PostPublishedJuly 18, 2019, 12:55 p.m....approaches: single-cell RNA-sequencing to identify neuronal class and deep sequencing of neurons from each class...of a GAL-4 driver that can bind to activation sequences upstream of genes to stimulate expression (Wang... -
Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells
TypeBlog PostPublishedSept. 26, 2019, 12:50 p.m....in place of SpCas9. gRNA A gRNA targeting a DNA sequence of interest is expressed in 293T cells and loaded... HDR allows for the precise insertion of a DNA sequence ranging from a single nucleotide change to large...utilize HDR, a donor template containing the desired sequence flank by homology arms must be delivered to cells... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog PostPublishedAug. 30, 2016, 2:30 p.m.... RNA sensors that can detect virtually any RNA sequence (Green et al, 2014). The second was an in vitro...laboratory. The detection threshold for these RNA sequences (3 nM), however, was quite high and above what...and prototyping pipeline that can take pathogen sequence information from public databases to the production...