...Microbial Opsins Opsins are light-gated ion channels or pumps that absorb light at specific wavelengths. ...respond to different wavelengths of light, like blue or yellow light. These various opsins also initiate...also a blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance...Channelrhodopsin from Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum... 566 ArchT Light-activated outward proton pump from Halorubrum strain TP009. Improved light sensitivity...binding domain, allowing light to activate transcription. Phytochromes and light oxygen voltage (LOV) domains...) BLUF domains bPAC Light-activated adenylyl cyclase from Beggiatoa sp. for light-induced cAMP modulation...

...the Fab fragments contain highly variable regions, shown in light blue and light orange, that bind to a ...are light-sensitive molecules which emit a specific wavelength of light after excitation by light of a...pair the brightness of the fluorophore with the abundance of the antibody target. If a bright fluorophore...molecular tools used in the lab to specifically and tightly bind to other proteins. Antibodies, produced as...-efficient way to produce antibodies, with the highest variability in antibodies produced. Cells - Antibodies...protein chains: two large heavy chains and two small light chains. These four domains come together to form... to be produced in a single individual, giving a high probability that an antibody will be able to recognize...

...joining (NHEJ) pathway and the less efficient but high-fidelity homology directed repair (HDR) pathway....also enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no ..., which are about the same size as SaCas9, with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum... downstream of the target site (termed left and right homology arms). The length of each homology arm ...available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...strands Prime editing and base editing both offer high efficiency and low indel formation compared to HDR...favor insertion in the T-RL direction, with the right homology end closest to the target site. Type V-...

...simplicity of production, and higher titers. However, the risk of RCA formation is higher for first-generation ...rAdV vectors are also referred to as "gutless" or high capacity AdV (HCAdV) vectors. Here, all viral sequences...adenoviral backbone plasmid have matching left and right homology arms which facilitate homologous recombination...the shuttle plasmid, but this method results in a higher background of non-recombinant adenoviral plasmids... used for rAdV vector construction, due to their high gene transfer efficiency. Other serotypes like Ad26...immunogenicity. For example, Ad5/35 and Ad5/26 combine the high transduction efficiency of the Ad5 serotype with... particle. Vaccine Development rAdV vectors are highly immunogenic because they activate both innate and...

...episome which can concatemerize, producing high molecular weight structures that are maintained extrachromosomally...hybrid capsid derived from eight different serotypes. AAV-DJ displays a higher transduction efficiency in...on transgene expression and the expression of neighboring genes. rAAV episomes can develop chromatin-like...in vitro than any wild-type serotype, and a very high infectivity across a broad range of cell types in...Brain Connectivity Atlas and the Janelia Campus MouseLight project . rAAV vectors have also been developed...signals of cells (commonly neurons) in response to light. The short coding sequences of channelrhodopsins...wild-type). Another split AAV method which has shown higher efficiency in delivering large genes makes use ...

...opto-chemogenetic fusion proteins of a light-emitting luciferase and a light-sensing optogenetic element. When...neurotransmission. This, and the fact that GPCR ligands can have high potency, which is potentially important for dosing...off-target effects in vivo and the receptors exhibited high levels of constitutive activity, making them less...luciferin) is added, the luciferase enzyme generates light that activates the opsin. The opsin may excite or...illumination), making LMOs a versatile option. Coupling brighter bioluminescent modules, such as luciferase-fluorescent...example, a study in rats using an hM4Di receptor might consider an alternative DREADD ligand such as DCZ... chemogenetics by using physical and biological light sources for opsin activation. Proc Natl Acad Sci...

...insert expression (i.e., a strong promoter directs high expression, whereas weaker promoters can direct ...determines the type of plasmid and dictates how it might be used in the lab. Below are some common plasmid...particles, that can infect your target cells at a high efficiency. Addgene's expanding viral service offers...Next Generation Sequencing (NGS) comes in. NGS is a high-throughput, multi parallel sequencing platform that... Plasmids 101: 5 factors to help you choose the right cloning method Plasmids 101: Restriction Cloning...

...will have the highest specificity. To increase specificity, you can also consider using a high-fidelity Cas...gRNA targeting 5′ exon or essential protein domains High-fidelity Cas enzymes increase specificity. Dual-...a mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9 and gRNA...

...choosing the right cloning method for your research project . The following guide will highlight several of...

...for fast and efficient generation of large-scale, high-titer preps. This is thanks to established retroviral... on a case-by-case basis. Retroviruses carry a slightly elevated risk of integrating into proto-oncogenes...clinical-grade gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing...

...immune system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based...technologies developed for optogenetics research utilize light to detect, measure, and control molecular signals...

...J., Bae, S., & Hur, J. K. (2021). CReVIS-Seq: A highly accurate and multiplexable method for genome-wide...Kochenderfer, J. N., Norberg, S. M., Hinrichs, C., Highfill, S. L., Somerville, R. P., Panch, S. R., Jin, ...

...additional hypothesis-directed experiments. As you might have guessed from their name, pooled libraries are...

...polymerase) to bind to a promoter. Transcription is tightly controlled in both bacteria and eukaryotes. Promoters...