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Experiment Tags: CRISPR, Genome Engineering, Cardiac Gene Editing, rAAV Delivery, Dystrophin Restoration
Experiment Summary: This protocol describes how to use the clustered, regularly interspaced, short, palindromic repeat (CRISPR) system to perform postnatal cardiac gene editing in mdx/utrophin+/- mice. Specifically, the protocol uses the Staphylococcus aureus CRISPR-associated protein 9 (SaCas9) and guide RNAs (gRNAs) delivered via recombinant adeno-associated virus (rAAV) serotype rh.74 (rAAVrh.74) to carry out the gene editing. Restoring dystrophin expression in these mice's cardiac muscles is the main goal in hopes of developing a treatment for Duchenne muscular dystrophy (DMD).
Experiment Tags: CRISPR/Cas9, Lentivirus, Gene Knockout, Cell Line Development
Experiment Summary: The protocol utilizes lentiviral vectors and a control system to regulate Cas9 activity, enabling the development of knockout cell lines. The development of a human muscle cell line that targets the gene serves as evidence of the protocol's effectiveness.
Experiment Tags: Gene Editing, FLPe Recombinase, Cassette Exchange, AAVS1 Locus, Human Pluripotent Stem Cells
Experiment Summary: The protocol presents a unique approach to FLPe recombinase-mediated cassette exchange (RMCE) gene editing at the AAVS1 locus in human pluripotent stem cells (hPSCs) that is both rapid and efficient. This method makes it easier to generate highly reliable isogenic lines, which in turn makes transgenesis studies in hPSCs easier to conduct.
Experiment Tags: CRISPR-Cas9, Gene Editing, C. elegans, sgRNA, Plasmid Construction Homologous Recombination, Genetic Screening.
Experiment Summary: This protocol outlines the process for performing CRISPR/Cas9-mediated genome modifications in C. elegans, including the design of specific sgRNA sequences, construction of Cas9-sgRNA plasmids, and preparation of injection mixtures.
Experiment Tags: Bacterial Transformation, Bacterial Transformation Protocol, transformation, bacterial cells
Experiment Summary: Bacterial transformation is a fundamental process in molecular biology, enabling the introduction of foreign DNA into bacterial cells. Primarily used for the amplification and storage of plasmids, bacterial transformation is critical for cloning experiments.
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