CRISPR Genome Editing
Clustered regularly interspaced short palindromic repeats, more colloquially known as CRISPR, are a genetic component of bacterial and archaeal antiviral immunity. They are composed of a series of palindromic repeats and spacer sequences. Spacer sequences are derived from the genomes of bacteriophages that have invaded the cell before making them a type of adaptive immunity. Many proteins related to this immune system are commonly referred to as CRISPR-associated (Cas) proteins and they allow the cell to generate and use spacer sequences.
Different Cas endonucleases and guide RNA (gRNA) form the core of CRISPR-Cas genome editing. A gRNA is composed of a scaffold sequence, which allows it to bind to a specific Cas protein, and an artificial spacer homologous to the sequence of interest. One of the most commonly used Cas proteins is Cas9 endonuclease: when bound to any gRNA strand it locates a homologous sequence and cuts both of the strands at a relatively specific location.
CRISPR Gene Knock Out
In CRISPR-Cas mediated knockout, an active Cas endonuclease (e.g. Cas9) and a gRNA designed to target the sequence of interest are introduced to a cell. Cas9 will eventually cut at the homology sequence and the cell will attempt to repair the damage done. The repair mechanism eukaryotic cells employ are not always accurate, and some small indels or SNPs can form at the cutting site. If these changes cause a frameshift or a nonsense stop codon, the gene product cannot be expressed.
Let Genome Editing Core help you with your lentivirus-mediated CRISPR knockout project from the very beginning to the end.
- Plasmid cloning and gRNA design
- Generation of lentiviral vector particles
- Transduction and KO population selection
- Generation of a clonal KO population by single-cell sort
Please, do not hesitate to contact Genome Editing Core for consultations on topics listed above or general study design.
CRISPR Genome Engineering
We also offer consultations and/or services related to other CRISPR/Cas methods such as:
- Lentiviral library screens
- Knock-ins and base editing
- Epigenetic modification by recombinant deactivated Cas proteins
Contact Genome Editing Core staff for more information on how we can enable your imagination!