We narrowed to 402 results for: MUT
-
TypeBlog Post...the preferred pathway for generating knock-in mutations, and NHEJ is an error-prone pathway frequently...frequently introducing frameshift knockout mutations. Thus, if genetic manipulation one way or the other (knock-in...
-
Multiplexed Overexpression of Regulatory Factors (MORF) collection
TypeBlog Post...example, very few diseases manifest as a single mutation or expression dysregulation event. Diseases are...multiple layers of dysregulation from either genetic mutation and/or TF dysregulation. A targeted TF screening... -
Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity
TypeBlog Post...Joung and Jennifer Doudna Labs used targeted mutagenesis to generate hyper accurate Cas9 (HypaCas9). This... sites. Using a combined random and directed mutagenesis approach, the Anna Cereseto Lab created pX-evoCas9... -
Plasmids 101: Screening Strategies Used in Plasmid Cloning
TypeBlog Post...non-functional 𝛽-galactosidase. Complementing this mutation by expressing this deleted portion, for example...custom primers are especially helpful in verifying mutations that wouldn't be reached by sequencing from either... -
Hot Plasmids - May 2022
TypeBlog Post...long sequences with reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally...traditionally have higher risks of off-target mutations. These plasmids contain catalytically inactive dCas9 and... -
GPCRs: How Do They Work and How Do We Study Them?
TypeBlog Post...chemogenetics receptors, GPCRs were first mutated via site-directed mutagenesis to bind non natural ligands. These... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog Post...further tested the base editors by correcting a mutation that causes Niemann-Pick disease type C, directly...McCormick lab generated knock-out and knock-in mutants, determined protein expression levels from different... -
Viral Vectors 101: An Introduction to AAV
TypeBlog Post...vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) to form ...genome with wild-type ITRs at the both ends and mutated ITR at the center of symmetry. After uncoating ... -
Transferable Skills Guide: Public Speaking
TypeBlog Post... You’re muted on Zoom. Your video conferencing software freezes and then crashes. You’re muted on Zoom... -
Plasmids 101: CcdB - The Toxic Key to Efficient Cloning
TypeBlog Post...gene. One such strain is DB3.1, which contains a mutant version of DNA gyrase (gyrA462) that is resistant... al. “F plasmid CcdB killer protein: ccdB gene mutants coding for non-cytotoxic proteins which retain ... -
Rabies and Neuronal Tracing
TypeBlog Post...with a deletion-mutant rabies virus. Retrograde neuronal tracing with a deletion-mutant rabies virus. Pubmed... -
Pathways Over Time Plasmids Engage Students in Functional Genomics Research
TypeBlog Post...proteins have acquired divergent functions due to mutations accumulated over time? Undergraduates can help...because the S. cerevisiae deletion strains are ∆ura3 mutants and cannot grow without uracil supplementation ... -
Design Tips for Prime Editing
TypeBlog Post...with BioRender.com. Add silent mutations near point mutations to create 3-base (or longer) tracts... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog Post...additional SapTrap plasmids used for introducing point mutations that was published in a microPublication (Schwartz... E. M. "SapTrap vectors for introducing point mutations with unc- 119 + selection." MicroPublication 1... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog Post...orientation limiting FRET efficiency, circular permutation (Baird et al., 1999) of the fluorescent protein...References 1. Baird, Geoffrey S., et al. "Circular permutation and receptor insertion within green fluorescent... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog Post...indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as well as any...functional consequences of that change. For example, “Mutated aspartate 123 to Lysine (D123K), lowers biosensor... -
Progress Towards a PAM-Free CRISPR
TypeBlog Post... if you want to knock out a gene, a frameshift mutation introduced anywhere within the early coding region... hand, if you are trying to knock-in a patient mutation or need to make a precise amino acid change, there... -
CRISPR in the Clinic
TypeBlog Post...co-injection of Cas9 and a single-guide RNA targeting the mutant allele. When all was said and done, the researchers...targeted to precisely edit with the goal of ‘fixing’ mutations or introducing a desired edit, but this particular... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog Post...genes with many modifications. We look for point mutations, truncations, and insertions that could compromise...compromise function. When we do find mutations, we check to see if they affect the translated amino acid. ... -
7 Simple Strategies to Resolve Conflicts with Difficult Supervisors
TypeBlog Post...your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor might not... his or her viewpoint, and discuss options for mutually beneficial solutions. If you package your challenges... -
Chromoproteins: Colorful Proteins For Molecular Biology Experiments
TypeBlog Post...promoter. The resolution step leads to the desired mutation and excision of the vector backbone. To do this...Characterization of a Blue Chromoprotein and Its Yellow Mutant from the Sea AnemoneCnidopus Japonicus. Journal... -
Plasmids 101: Optimizing Plasmid Yields
TypeBlog Post...due to its endA1 recA1 relA1 genotype, as these mutations improve plasmid stability and yield. If a plasmid...using older plates could increase plasmid loss or mutation. Let it incubate Increasing the incubation time... -
Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards
TypeBlog Post... E. The mutagenic chain reaction: A method for converting heterozygous to homozygous mutations. Science... -
Tips for Using BLAST to Verify Plasmids
TypeBlog Post...BLAST (blastx) search to check for expected ORFs, mutations or truncations. A primary advantage of blastx ...database. Another benefit is that a frameshift mutation present in the ORF is readily apparent when viewing... -
Which Fluorescent Protein Should I Use?
TypeBlog Post...outlined below (or emission wavelenght range). By mutating GFP, the variants blue FP (BFP), cyan FP (CFP)...breakdown of GFP, it's variants, and their relevant mutations, check out Marcy's post from last week on GFP.... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog Post...which contains multiple point mutations in the nuclease domain. These mutations result in an inactive RNA-... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...et al. 2018). This was accomplished using a Cas9 mutant (Cas9M1C/C80S) that harbors two surface exposed...disulfide-activated ligands. Due to the concerns of off-target mutations and transgene integration in crops, delivery of... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...success of BiFC experiments. The ideal control is a mutated variant of one or both of your PIPs in which the...requires that you empirically test a number of mutants to identify those that inhibit BiFC in your cellular... -
Plasmids 101: Colony PCR
TypeBlog Post... but won’t tell you if your products have any mutations. There are lots of different cloning strategies...expected sized PCR product doesn’t mean there aren’t mutations in your insert. Make sure to submit multiple positive... -
Your Lentiviral Plasmid FAQs Answered
TypeBlog Post...the deletion reduces the risk of insertional mutagenesis by limiting the interaction between viral LTR...potential for oncogenesis through insertional mutagenesis. These risks are largely based upon the vector... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post... the sequence, normally through the synonymous mutation of a codon. You can domesticate the sites yourself...colony PCR electrophoresis), is free of undesired mutations, and that you have the correct four-base overhangs... -
Hot Plasmids Spring 2024
TypeBlog Post...HaloTag, Figure 4), they determined that an A122D mutation in the rhodopsin domain conferred several new ...potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog Post...methods were available to create targeted gene mutations in anoles lizards or in any reptiles at all. But...was able to make an albino lizard with targeted mutations. Now what? What would you study first? They are... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog Post...determine the presence and quantity of DNA, RNA or a mutation of interest. Together SHERLOCK and DETECTR demonstrate...SHERLOCK could also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog Post... HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a molecule results in...Zika virus, genotype human DNA, identify tumor mutations within cell-free genomic DNA, and detect COVID... -
Mesothelioma - Causes, Symptoms, and Treatment
TypeBlog Post...inflammation, which can cause mesothelial cells to mutate and ultimately form cancerous tumors. This process...For example, multiple studies show that certain mutations of the BAP1 gene play a role in susceptibility... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog Post...mEGFP. This variant was made by introducing two mutations (F64L, S65T) in the original jellyfish GFP (AvGFP...with improved brightness (Tsien, 1998). Another mutation (A206K) is necessary to generate the strictly ... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog Post...biosensors are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...around 70 mM and 100 mM, respectively. Introducing mutations and using potassium-binding protein homologues... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog Post...2013) Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas...Rice Blast Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog Post... can mediate recombination.) The deletions are mutually exclusive, as either deletion removes one of the...following the promoter. The STOP cassette includes a mutant YFP that does not fluoresce, but can be detected... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog Post... spiroplasma (M. Sss1) (termed MQ1.) The Q147L mutation improves methylation kinetics such that cytosine...fused LSD1 to x-Cas9(3.7). x-Cas9(3.7) is a SpCas9 mutant that exhibits increased PAM flexibility, and this... -
9 tips for a successful postdoctoral experience
TypeBlog Post...with senior investigators. Discussions can reveal mutual scientific interests, complementary perspectives...research to search committees, and can lead to mutually beneficial collaborations. 7. You cannot get too... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog Post... into question. Specifically, CRISPR-generated mutants believed to be knockouts can maintain partial function...process is tedious and time consuming, as RNAi mutants with a reduced growth phenotype will require additional... -
Important Considerations When Using AAVs
TypeBlog Post...them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses... Russell, David W. "AAV vectors, insertional mutagenesis, and cancer." Molecular Therapy 15.10 (2007):... -
Quick Guide to All Things Lentivirus
TypeBlog Post...active area of research due to the possibility that mutation or recombination with human retroviruses could...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog Post... most often consist of an enzyme that has been mutated to block its catalytic activity and instead become... later work showed that introducing a Leu28Cys mutation in eDHFR does lead to covalent binding of the ... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog Post...especially for G-protein-coupled receptors (GPCRs). Mutations in receptor sites can affect the fluorescence ...doi.org/10.1038/s41592-021-01338-6 Grimm, J. B., Muthusamy, A. K., Liang, Y., Brown, T. A., Lemon, W. C.,... -
Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart
TypeBlog Post...creation of improved fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent... -
CRISPR Cheat Sheet
TypeBlog Post...multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of interest. ... -
New Podcast Segment: Hot Plasmids
TypeBlog Post... new tool for making diverse, localized point mutations. Listen to Our Segment on CRISPR X Hot Plasmid...