Sgrna Construction
Sgrna Construction - Related Videos
02:15
Nanoblade-Based Delivery of Nucleic Acid Cargo: A Technique to Deliver Cas9-sgRNA Complex to Target Cells via Virus-Like Particles
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2025
This video demonstrates a technique for nanoblade-based delivery of Cas9-sgRNA ribonucleoprotein complex inside target cells for genome editing. Nanoblades are virus-like particles devoid of the ability to multiply and infect neighboring cells, therefore are very useful for rapid and dose-dependent transportation of bio- and nanomaterials.
13:43
Fabrication of Myogenic Engineered Tissue Constructs
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Cited by 11 •
2009
Here, we demonstrate fabrication of collagen-based, tissue constructs containing skeletal myoblasts. These 3-D engineered constructs may be used to replace or repair tissues in vivo. For our purposes, we have designed these as an atrioventricular electrical conduit for the repair of complete heart block[1].
13:48
Design and Construction of an Urban Runoff Research Facility
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Cited by 6 •
2014
This paper describes the design, construction, and function of a 1,000 m2 facility containing 24 individual 33.6 m2 field plots equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated home lawns.
10:08
Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests
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Cited by 4 •
2017
This paper describes the assembly process and operation of a bench-scale photosynthetic bioreactor that can be used, in conjunction with other methods, to estimate pertinent kinetic growth parameters. This system continuously monitors the pH, light, and temperature using sensors, a data acquisition and control unit, and open-source data acquisition software.
10:10
HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
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Cited by 3 •
2019
A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.
FAQs
The target-complementary sequence provides the nucleotide-level recognition information, while the structural scaffold preserves the RNA architecture required for association with Cas9. These components must function together: recognition alone does not provide the enzyme-binding structure, and the scaffold alone does not specify a genomic location. Their coordinated design allows the assembled guide to direct Cas9 toward the intended DNA sequence.
A suitable protospacer adjacent motif, or PAM, must occur next to the selected target region for Cas9-mediated recognition and cleavage. Consequently, target selection cannot rely only on sequence complementarity. Designers must evaluate the target and its neighboring DNA context together, because an otherwise attractive complementary sequence may not support the positioning required for enzyme activity.
Careful guide design links target selection with the correct complementary sequence, structural scaffold, and nearby PAM. This coordinated selection supports recognition of the intended nucleic acid region rather than relying on an incomplete guide configuration. Consistent construction also improves experimental reproducibility, allowing different bioengineering experiments to use guides that are assembled and evaluated according to the same design logic.
A typical workflow begins by selecting the nucleic acid region to be addressed, identifying an appropriate adjacent PAM, and designing the target-complementary sequence. That sequence is then joined with the structural scaffold derived from CRISPR RNA and trans-activating CRISPR RNA. The resulting guide is prepared for Cas9 binding and evaluated in relation to the intended editing objective.
These applications use guide design to direct Cas9 activity toward a selected genomic region. For gene knockout, the target is chosen to disrupt the gene, whereas sequence insertion uses a target site suited to introducing new sequence information. In both cases, the guide must connect the intended locus with the enzyme’s recognition and cleavage requirements.
The same design principles can be adapted beyond permanent sequence changes. Guides directed to selected regions can support transcriptional regulation, while collections of guides can enable genome-wide screening across many targets. These uses make guide construction important in bioengineering because the quality and consistency of individual guides influence how reliably researchers interrogate genes or regulatory regions at scale.