Protocol Generator from knock in to WB-Gene Editing Protocol Design

Example

CRISPR/Cas9 mediated homologous recombination is employed to insert a 2A-GFP reporter before the stop codon of the MYF5 gene in human iPS cells. This allows for the prospective identification and purification of myogenic progenitors.

Scenario

Researchers can use this function to study early myogenic specification or other cellular differentiation processes by tracking the expression of the reporter gene integrated into a specific genetic locus.

Example

Following the generation of knock-in reporter lines, the Protocol Generator guides users through the steps for characterizing the genomic modification. This includes PCR and sequencing validation of reporter clones to confirm in-frame positioning and absence of off-target effects.

Scenario

This is particularly useful for validating the success of the genetic modification and ensuring the reliability of the experimental model before proceeding with further experiments.

Example

After genetic modification and cellular differentiation, the Protocol Generator provides detailed steps for protein extraction, gel electrophoresis, and immunoblotting to detect the expression of target proteins, such as the ASC oligomerization in macrophages.

Scenario

This function is crucial for researchers aiming to verify the protein-level effects of genetic modifications, such as the expression of a reporter protein or the impact of gene editing on protein interactions and pathways.