We narrowed to 815 results for: MAL
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TypeBlog Post...create SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects...Another way to improve base editing efficiency for mammalian edits is to ensure the editors make it into the... CBEs (called Td-CBEs, CBE-Ts, or TadCBEs) are smaller and less likely to cause off-target mutations than...2023). Evolution of an adenine base editor into a small, efficient cytosine base editor with low off-target...
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CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...efficiency for point mutations and 66% efficiency for small insertions. Multiplexing of two targets simultaneously.... coli; while the Marraffini lab used a native minimal CRISPR array (Bikard et al., 2013), the Qi lab ... plasmids Stanley Qi’s lab demonstrated that a minimal, regulatable CRISPR system effectively silenced...transcription of one or more genes in E. coli. and in a mammalian cell line (Qi et al., 2013). Two important features... -
Sensing Neuronal Dopamine
TypeBlog Post...method is that it can be performed in a live awake animal by surgical implantation of the probe into a specific...powerful and offers immediate measurements in a live animal that can be correlated to behavior, the technique... molecular tool to detect and image DA in live animals (Patriarchi et al., 2018). Similar to calcium and... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post...specialized Cas proteins snip the foreign DNA into small fragments approximately 30bp in length and paste...editing was initially described by Cong et al. and Mali et al. in January 2013 (7, 8). Nine months later...Zhang F. Science. 2013 Jan; 339 (6121): 819–23. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE... -
Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource
TypeBlog Post...Polyclonal antibodies produced in animals may be variable among animals and bleed dates. The development...scientific community. Why recombinant antibodies? Animal-based methods for antibody production have several... -
What Do I Do Now? Academic v. Non-Academic Career Decisions
TypeBlog Post... Step Further - Big Company v. Small Company Big (e.g. Pharma) Small (e.g. Biotech) Develop expertise...ride Complicated matrix structures are common Small integrated teams, little hierarchy Real diversity... -
10 Great Guest Posts We're Thankful For
TypeBlog Post... experiments. Better Dyeing Through Chemistry & Small Molecule Fluorophores In this post, Luke Lavis from... A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are commonly used... MIT) shows how you can use CombiGEM to barcode small libraries of CRISPR gRNAs to use for screening experiments... -
Learning Technical Skills at a Lab Central Workshop
TypeBlog Post...a DNA sample into many small components. Without proper processing, these small DNA strands can form concatemers... produced high yields of quality libraries from small amounts of DNA i.e 100 pg. At the start of the workshop... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog Post...was able to translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, ...with light Article contributed by Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule ...loops to create genomic interactions that wouldn’t normally occur. The synthetic looping factors developed... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog Post...based multicolor labeling system to visualize chromosomal loci in human cells (Ma et al., 2015). To do ...aptamers are nucleic acid sequences that bind to the small molecule, DFHBI-1T to induce its fluorescence. They...Hanhui et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proceedings of the National... -
DAP arrays for multiplex gene editing
TypeBlog Post...MBE) and multiplex prime editing (MPE) with only minimal payloads. DAP array Although CRISPR-Cas12a, with...advantages in gene editing, while keeping the system to minimal size. To do so, they harnessed the tRNA processing...any cell type or organisms of interest with only minimal genetic payloads. Different from existing multiplex... -
Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**
TypeBlog Post...portion of the complex and is derived from the malachite green (MG) dye, but has been modified with the...et al also show that MG-2I-dL5** only generates small amounts of non-specific ROS when exposed to YFP ...Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on Addgene.org ... -
Science Career Options
TypeBlog Post... opportunity to advance, and how many jobs are normally available (as sectors such as forensics currently...the relevant skills and experience.You might not normally think about it, but hobbies and volunteer experiences...are willing to exercise these skills outside of a formal work environment also shows that you are passionate... -
Synthetic Photobiology: Optogenetics for E. coli
TypeBlog Post... biological processes. Chemical effectors are suboptimal for a number of reasons, including their potential...number of plasmids, with one RR (ompR) encoded chromosomally. Leakiness was also an issue; some promoter ...transcription factors, for which low expression levels are optimal. These tools are compatible with many strains of... -
Choosing Your Fluorescent Proteins for Multi-Color Imaging
TypeBlog Post...supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved folding...however, that there is no single fluorescent protein optimal for all organisms and that if you want the brightest... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e., 10 and 50 µL of vector at 1 x 1013 vg/mL... Hudry et al, 2023; Hinderer et al, 2018). The optimal dose for each particular experiment is determined...validate the capsid for your specific application and animal model. Other model organisms Tropism and efficacy...transduces BALB/cJ, CBA/J, and FVB/NJ mice, with minimal liver transduction observed in both BALB/cJ and...and AAV-PHP.S, with a bias toward segments of the small intestine (Chen et al., 2022). Systemic capsid for... -
Viral Vectors 101: Gamma-Retroviral Packaging Systems
TypeBlog Post...envelope, Phoenix-AMPHO (for the infection of mammalian cells.) Using this system, virus is produced in...the host cell cytoplasm, the viral components are small enough to enter the nucleus through the nuclear ...of retroviruses for efficient gene delivery to mammalian cells using 293T cell-based systems. Curr Protoc... -
Viral Vectors 101: Viruses as Biological Tools
TypeBlog Post... There are four main types of viruses used in mammalian research: 𝛄-retrovirus, lentivirus, adenovirus...Mutagenesis Possible Adenovirus Remains Episomal (Little Risk of Insertional Mutagenesis) High ...Expression High Immunogenecity AAV Remains Episomal (Little Risk of Mutagenesis) Very Low Immunogenicity... -
Plasmids 101: Screens vs. Selections
TypeBlog Post...or mutagenizing a population (typically cells or small organisms) and then assaying for a functional or...are genetic knockout mutations and treatment with small molecules/drugs. With the advent of CRISPR, genome-wide...be knocked out with CRISPR, the entirety of the small molecule realm is typically not feasible to test... -
Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection
TypeBlog Post...infected bacterium can either exist as transient extrachromosomal DNA, like a plasmid, or it can integrate into...introduce foreign DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral and...chromosome and allows the donor bacterium to make a small “bridge” or sex pilus that attaches to the recipient...