CRISPR-Cas9 is a popular method that brings researchers endless experimental strategies to create their own research-based cellular models. In this post we’ll review a new transfection reagent especially engineered to maximize Cas9 vectors deliveries inside cells with low cellular toxicity.
Cas9 mRNA optimized for genome editing
CRISPR/Cas9 is relatively simple to implement, as the researcher fully controls the experimental design of the tools, from the sgRNAÂ sequence to the Cas9 protein.
Let’s quickly look back to the basics… CRISPR Associated Protein 9 (Cas9) functions as part of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) genome editing system.
In this system, a single guide RNA (sgRNA) sequence targets a site of interest in the genome while the Cas9 protein performs a double stranded DNA cleavage at the targeted genomic location. CRISPR/Cas9 system can be widely used to express or silence a gene, create mutations and variants… into the genome of the host cell.
All GeneCopoeia products and services available from tebu-bio
GeneCopoeia brand products include any possible DNA construct/plasmid in addition to an ever-growing offer of associated products and services. Here is a quick overview of the offer:
If you search for any human or murine gene (e.g. NFKB1) on the GeneCopoeia website, you will get a search result like this one displaying the available catalog products: [Read more…]
CRISPR genome editing: which cell line to choose?
Many labs have adopted the CRISPR genome editing technology to make knock-out and knock-in cell lines.
This technology produces first a targeted break in genomic DNA, which can then be exploited to produce cell lines with genes knocked out or where a donor vector has been used to introduce new genetic elements (point mutants, fluorescent tags, antibiotic resistance cassettes, etc.). Essentially any desired modification to the cells genome can be made. In setting up these genome editing projects there are many choices to be made including vector for the Cas9 protein and for the sgRNAs. Perhaps the most difficult choice, however, can be which cell line to use. Even the most affordable stable genome editing cell line development services can come with a significant cost, so choosing the right cell line at the beginning is crucial. Here we explain some of the choices researchers have in setting up their CRISPR genome editing projects and give our advice for cell line selection.
Using CRISPR to knockout an essential gene
Genome editing technology enabled by CRISPR and TALEN has become mainstream. Most cell biology labs are engaged in projects to create custom cell lines with knock-outs and knock-ins, and companies such as GeneCopoeia even propose complete cell line generation services. Projects can involve transfection of mammalian expression constructs, TALEN pairs, or direct transfection of RNA.
When scientists want to make a stable knockout cell line, one of the first questions they should ask is whether or not the resulting cell line will be viable. Often a murine knockout mouse has been made and/or siRNA knockdown experiments have been performed in human cells, so experienced users have a good idea if a human knockout cell line will be viable. There are certainly some cases, however, where either the researcher knows or expects that a complete knockout will not be viable but wishes to make the knockout nonetheless. What is the best way to deal with all of the risk involved with starting an relatively expensive and time-consuming project like this that could end in failure? [Read more…]
Genome Editing in Stem Cells: outsource or do-it-yourself?
Many researchers are facing a dilemma: they want to set up a CRISPR genome editing project but they can’t decide which cell line to use for genome editing. Even some of the most cost-effective genome editing cell line generation services like the one from GeneCopoeia will cost a few thousand euros, so picking the correct cell line and setting up the project correctly is very important. Researchers working in primary cells may find the idea of switching to an immortalized cell line a bit artificial. They dream of the possibility of editing the genome of a stem cell line which they can then differentiate into their tissue of choice as needed. [Read more…]
Genome-Editing by Mail
Due to the development of CRISPR and TALEN technologies, the field of genome editing is evolving rapidly.  This post highlights the natural evolution of these technologies leading to outsourcing gene editing to skilled service providers.
What's Trending in the Molecular Biology World? CRISPR !!!
It’s official, CRISPR has officially reached “Breakout” status as a Google search term (see image from Google Trends). The Clustered, Regularly Interspaced, Short Palindromic Repeats (CRISPR) and RNA-guided Cas9 nucleases are an exciting new gene editing tool that allows life science researchers to directly modify the DNA of the organism they study.