We narrowed to 316 results for: Mut
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TypeBlog PostPublishedMay 7, 2020, 1:15 p.m....efficient. Informed by these mutations and previously described TadA mutations (Gaudelli et al., 2017), they...withstand antibiotic selection. As a result, 8 mutations in TadA could create all three edits and were ...placeholder for the number that identifies the mutation in the evolved TadA domain. Because ABE7.10 contains...therapy approaches can use base editors to correct mutations in stem cells and return the corrected cells back...tested ABE8.17-m and found that one base change mutation (ABE8.17-m+V106W) (Gaudelli et al., 2020) was ...
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Hot Plasmids and Viral Preps - May 2021
TypeBlog PostPublishedMay 18, 2021, 1:15 p.m....preps Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz As multiple antibody-based...coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom’s lab... library encompassing all possible amino acid mutations of the receptor binding domain (RBD) of the viral...et al Cell., 2020. (B) Mapping antibody-escape mutations with the yeast display library. From Starr et ...antibodies. In the library, each plasmid encodes the mutated spike protein and a barcode, and is designed so...The Bloom lab used the library to identify RBD mutations which do not interfere with ACE2 binding, but ... -
Delivery Methods for Generating iPSCs
TypeBlog PostPublishedApril 17, 2018, 1:37 p.m....Efficient and Stable Genome integration Insertional mutagenesis, transgene reactivation, residual expression ...Efficient and Stable Genome integration Insertional mutagenesis, residual expression piggyBac Excisible Medium...sequence cells to verify excision didn’t introduce mutations Adenovirus Non-integrating Low Medium Generates...generating methods due to the risk of insertional mutagenesis. Random integration also creates heterogeneous...Finally, retroviruses are prone to insertional mutagenesis and random integration creates variation between...because they are less likely to cause insertional mutagenesis, adenoviral vectors are considered a safer way...These methods eliminate the risk of insertional mutagenesis, the presence of a genetic scar, and incomplete... -
Adeno-associated Viruses (AAVs) for Genome Editing
TypeBlog PostPublishedMarch 27, 2018, 1:32 p.m.... introduce naturally occurring cancer-causing mutations into the endogenous allele of the STAG2 tumor ... introducing non-sequence specific frameshift mutations through aberrant double strand break repair, it...epitope tags, or the introduction of dominant mutations of a gene into the endogenous allele in cultured...template to mouse zygotes and thereby correct a mutation that causes Hemophilia B. Unfortunately, work ...segregation in human cells harboring tumor-derived mutations in STAG2." PLoS genetics 12.2 (2016): e1005865...using adeno-associated virus (AAV)." Chromosomal Mutagenesis. Humana Press, New York, NY, 2015. 75-103. PubMed... -
Evolution of Lab Techniques
TypeBlog PostPublishedJune 21, 2016, 2:30 p.m....particular gene mutation, southern blotting can be used to identify the presence of that mutated gene. In southern...the presence of the mutated β globin gene that causes sickle cell anemia. The mutation happens to destroy...the probe (for example, has the sequence of the mutated gene the researcher is searching for), then the...southern blotting being used to identify a gene mutation is the genetic test used for sickle cell anemia... -
Editor's Choice, August 2016
TypeBlog PostPublishedSept. 2, 2016, 1:32 p.m....optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...personal experience, is a quick way to generate many mutated plasmid constructs. The second post was contributed...specificity issues, and other means of generating mutations in live cells are all discussed further in this...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course... in chronological order here: Site Directed Mutagenesis by PCR Google Forums Round Up: First Impressions... -
CRISPR 101: Drug discovery and CRISPR
TypeBlog PostPublishedMay 4, 2023, 1 p.m....knock-ins of patient mutations which are variants of unknown significance – mutations with unclear effects...with defined, straightforward causes (a specific mutation in a known gene = disease onset) may be best served...accessible, fast, and easy. In cases where the mutational basis of a disease is known – repeat expansion...CRISPR could even be used to generate patient mutations and then test both individual and combinatorial... -
CASTing Off for New Shores in Human Genome Editing
TypeBlog PostPublishedJune 10, 2025, 1:15 p.m....over (with a little mutagenic assistance). Quicker CAST activity allowed certain mutated phages to outcompete...Witte et al., 2025). The best evoCAST featured 20 mutations in TnsB, the subunit that catalyzes DNA insertion...therapy, highly tailored treatments to specific mutations require individualized optimizations, safety testing...intervention for a variety of loss-of-function mutants could provide a streamlined, one-size-fits-most... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog PostPublishedNov. 1, 2016, 2:30 p.m....several struggles with these techniques: unwanted mutations due to excessive PCR cycles or low fidelity Taq...purify and sequence the plasmid to confirm no mutations were introduced during PCR or any subsequent steps...fidelity DNA polymerases, I have never had any mutations introduced into my inserts. However, with any ...technique there is a chance of introducing an unwanted mutation. Tips for high efficiency cloning Now that you...Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your Plasmid Background with... -
Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS
TypeBlog PostPublishedJuly 21, 2017, 12:48 p.m....there might be a mutation that disrupts the function of YFG. You could compare this mutant fly to one lacking...lacking the mutation in the same genetic background. In other cases, YFG or one of its mutant variants will... knock down gene expression, rescue knock out mutations, and overexpress viral genes. Additionally, other... -
Viral Vectors 101: Viruses as Biological Tools
TypeBlog PostPublishedMarch 21, 2023, 1:15 p.m....the fear of random genomic integration causing mutagenesis in the host. Adenoviruses can deliver relatively...Dividing Cells Moderate Immunogenicity Insertional Mutagenesis Possible Lentivirus Infects non-dividing...non-dividing cells Insertional Mutagenesis Possible Adenovirus Remains Episomal (Little Risk of...of Insertional Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity (... AAV Remains Episomal (Little Risk of Mutagenesis) Very Low Immunogenicity Infects Non-Dividing... -
Plasmids 101: Blue-white Screening
TypeBlog PostPublishedJune 4, 2015, 1:03 p.m....α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional... that deleting a section from the lacZ gene (a mutation called lacZΔM15) creates a non-functional β-galactosidase...will not complement the cell's β-galactosidase mutation. An unsuccessful cloning reaction leaves the α-peptide...proper E. coli strain (i.e. contains the lacZΔM15 mutation): XL1-Blue, DH5α, DH10B, JM109, STBL4, JM110, ... -
Plasmids 101: Yeast Vectors
TypeBlog PostPublishedFeb. 25, 2014, 7:11 p.m....limited to: rapid growth, ease of replica plating and mutant isolation, a well-defined genetic system, and a...recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids used in the lab typically...high rates of spontaneously occurring resistant mutants and the insensitivity of yeast strains to some ...missing nutrient. Scientists can exploit these host mutations by including a copy of a functional gene which... S. pombe no no *FOA in fission yeast induces mutation in the ura5+ gene in addition to ura4+. leu1... -
What's New in CRISPR - November 2021
TypeBlog PostPublishedNov. 18, 2021, 2:15 p.m....dominant negative mutation in MLH1 (PE4 = PE2+MLH1dn and PE5 = PE3+MLH1dn). Silent mutations added near the...the Cas12f scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI... and base editing, whereas a different set of mutations (the “V3.1” version) works best for genome editing...boosts the efficiency of prime editing to correct mutations in a variety of target genes and cell types. Their... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....Introduction of a conditional point mutation or functional mutation in vivo: Similar to a conditional rescue...fragment. The availability of both wild-type and mutant loxP sites has allowed scientists to leverage this...wild-type pair (loxP, black arrowheads) and one mutated pair (for example- lox511, white arrowheads). It...conditionally test the function of various point mutations and truncations in the absence of the endogenous... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m....and select for Cas9 mutants with PAM flexibility. In this system, the mutagenesis plasmid (MP6) permits... Base editing, another method to create point mutations, also has a strict editing window about 15 nt ...NGAG and NGCG and SaCas9 with an NNGRRN PAM by mutagenizing the PAM-interacting domain. However, these alternative...permits quick, large-scale mutation of the gene of interest. They created a selection phage (SP) to carry ... -
Polymerase Chain Reaction Overview and Applications
TypeBlog PostPublishedAug. 10, 2021, 1:15 p.m....colony PCR, or for generating mutated sequences for site-directed mutagenesis. Quantitative polymerase chain...sequences that are rare, for example, rare alleles or mutations. ddPCR also does not require a reference or standard...interest Molecular Cloning Site-directed mutagenesis Molecular Diagnostics qPCR Quantify starting... PCR Plasmids 101: Colony PCR Site Directed Mutagenesis by PCR Resources on Addgene.org Protocol: Polymerase... -
Simplify Cloning with in vivo Assembly
TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m.... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer... N-terminus of a protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag...rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase... -
22 Hot Plasmid Technologies from 2014
TypeBlog PostPublishedJan. 6, 2015, 5:21 p.m....Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab...mini-Tn5 vector that can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters...this vector, called pBAM1, by conducting random mutagenesis in the soil bacterium Pseudomonas putida and ...three dimensional structure of GFP, its brighter mutant S65T-GFP, and the subsequent biochemical characterization...proteins are derived from the Timer protein - a mutant of the fluorescent protein dsRed that changes irreversibly...recombination template vectors, to perform multiple site mutagenesis and create complex fusion or recombination vectors...The group further optimized Chrimson through mutagenesis (K176R) to improve the otherwise slow off-kinetics... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m....different mutations found in leukemia, the Lowe lab was able to identify a set of mutations that made ...Lowe’s lab utilized mice to study how different mutations in leukemia impact different treatment regimens...by a specific gene. For example, several genes mutated in human cancers have a homolog involved in yeast... yeast: A model organism for studying somatic mutations and cancer." Nature Education 1.1 (2008): 183....