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TypeBlog Post.... The last step involves a chemical reaction to stain the cells that contain this mRNA-riboprobe-antibody.../79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver, C.C. Mello. Potent and specific genetic...
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Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog Post...that treat disease at the molecular level. For instance, if a disease is caused by a mutation in a single...developing AAV-based therapies. Furthermore, deeper understanding of disease pathogenesis will enable robust therapeutic... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog Post...Create promoters with sequences upstream of the start codon of retinal cell-type specific genes from mice...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image... -
Plasmids for Endogenous Gene Tagging in Human Cells
TypeBlog Post...use electroporation, a technique that briefly destabilizes the cell’s outer membrane and allows the components...structures and processes and can lead to the better understanding of cell biology. We hope that these high-quality... -
Plasmids 101: FLEx Vectors
TypeBlog Post...considered "Cre-On" meaning that your gene of interest starts in the inverse/antisense "off" position and is ...employed for "Cre-off" (i.e. your gene of interest starts in the sense orientation and Cre is used to flip... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog Post...et al., 2024) have highlighted how chromatin substantially affects prime editing outcomes across many genomic...with PRIDICT/PRIDICT2.0, ePRIDICT can help you understand how chromatin might influence editing results... -
Fluorescent Tagging of Endogenous Genes with SapTrap
TypeBlog Post...light up endogenous genes of interest to better understand normal protein function. However, CRISPR protein...introduced directly into a Drosophila S2 cell line stably expressing Cas9. This PCR toolkit offers C- and... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...constructs are expressed transiently by transfection or stably impacts the success of BiFC experiments. In either...negative controls because their expression level, stability, and subcellular localization may differ from ... -
MXS Chaining
TypeBlog Post... (MCS). Modules are assembled one at a time by standard restriction enzyme digestion followed by ligation...Goler JA, Arkin AP, et al. BglBricks: A flexible standard for biological part assembly. Journal of biological... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog Post...CNS and PNS! What is the CREATE method? CREATE stands for Cre Recombinase-based AAV Targeted Evolution...number of neurons overall (GFP+ cells that also stained for a neuronal marker). Check out tables 1 and ... -
Simplify Cloning with in vivo Assembly
TypeBlog Post...is carefully designed primers. The majority of mistakes are due to incorrect primer sequences! Primers...transduction, and uses electron cryo-microscopy to understand receptor function. Follow them on Twitter... -
INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons
TypeBlog Post...randomly, or at fixed genomic target sites. This obstacle was recently surpassed with the discovery of CRISPR-transposons...genome engineering in bacteria, alongside more established tools such as Cas9-recombineering, CRISPRa/i,... -
Negotiating Work and Life: How to Find the Joy
TypeBlog Post... should not even be the real goal. We are all constantly negotiating our energy and attention. The goal...going away to a conference. Really. No one will starve while you are away a few days. Also, your children... -
CRISPR 101: Any Base Transversion Editors
TypeBlog Post...Tong, Wang, et al., 2023). Based on this promising start, the team used rational mutagenesis to produce the...narrowed the editing window and reduced A → G bystander edits. Their final tool, ACBE-Q (for an N108Q ... -
Hot Plasmids - August 2020
TypeBlog Post...fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed gRNAs and ...includes a CRISPRi-based synthetic oscillator, bistable network, stripe pattern-forming incoherent feed-forward... -
Using Video to Share Your Science: We Share Science
TypeBlog Post...tool for sharing ideas. This leads to everyday obstacles for many of us, such as finding time to read all...platforms (such as social networking, visual appeal, instant sharing across multiple other sites), I created... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...role as a receptor for PHP.B. The Deverman lab started by seeing how changes in LY6A’s expression affected... References Holmes, Christina, and William L. Stanford. "Concise review: stem cell antigen‐1: expression... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog Post... T’s. Thus, it’s important that there are no “bystander bases” if you choose this system. Fig....demonstrated that targeting dCas9 to transcriptional start sites was sufficient to repress transcription by... -
CRISPR Activation: A Practical Guide
TypeBlog Post...prevent your gene from being up-regulated. For instance, generally, the more highly expressed a gene is...whichever guides are closest to the transcription start site (TSS). We recommend targeting the guides to... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog Post...split vectors has been inconsistent. A greater understanding of basic AAV biology, however, would help improve...and effective in the retina of a mouse model of Stargardt disease. Hum Mol Genet 24:6811–6825 . https://...