This article is adapted from the original, which first appeared on The Bench, the Synthego blog.
CRISPR genome engineering is quickly becoming a standard laboratory technique due to its ease of use and precision. Synthego has increased the accessibility of CRISPR through its products and services, allowing more scientists to incorporate gene editing into their research.
With its increasing popularity, CRISPR technology has been applied to many different fields of research. Proteomics, for instance, traditionally focused on biochemical methods for isolating proteins and studying their interactions in in vitro assays. Now, researchers are using CRISPR to modulate protein profiles by editing genes and studying the downstream effects.
As more and more researchers are now trying their hand at CRISPR for the first time, products that simplify their journey are needed. Synthego has recently taken a bold step in this direction that will save CRISPR researchers months (or in some cases, years) of tedious lab work. The company now offers Engineered Cells products, including knockout cell pools, knockout cell clones, and advanced cells.
John LaCava, Research Assistant Professor at The Rockefeller University, NY, recently transitioned to using CRISPR in his research with Synthego’s Knockout Cell Pool product.
Dr. LaCava is a proteomics expert; his work focuses on developing technologies for proteome and interactome research. The general goal of his research is to better understand cellular biology by studying intermolecular protein interactions using approaches that allow structural and biochemical studies of purified protein complexes.
Recently, while working on transcription factor interaction networks, LaCava discovered that these networks are heterogeneously comprised of a collection of different complexes that are often mutually exclusive. In other words, different protein complexes may include one or more common components, but they don’t necessarily participate in the same biological functions.
This discovery led him to study how altering expression of a protein that is a component of multiple complexes (referred to as a nodal protein) affected the interactions of other proteins to better discriminate the physical entities that associate with each other.
That is how LaCava’s lab, that historically relied on biochemical methods for segregating out various complexes of interest, ended up going maverick with the CRISPR genome editing technology. Naturally, LaCava was concerned regarding successfully implementing the technique with the limited experience and infrastructure in his lab.
Dr. LaCava learned about Synthego’s Knockout Cell Pool product as a potential solution to get started using CRISPR is his research as quickly as possible, and when his colleagues validated the company’s product quality and customer service, LaCava decided to try it out.
Synthego’s Knockout Cell Pool products provide cell populations with the researcher-requested genes knocked out (>50% decrease in protein levels guaranteed) in the most relevant cell type for their research (more than 700 human cell lines are currently supported), allowing rapid generation of reliable data in their experiments.
“It is our first foray into CRISPR,” said LaCava. “Since we are novices, it can be intimidating to set up new assays. I thought Synthego’s Engineered Cells products presented us an opportunity to start out using CRISPR by only doing half of the work.”
LaCava aims to purify his target protein from the knockout cell pools and perform quantitative mass spectrometry to re-characterize the levels of proteins that have co-purified with the target protein. Significant changes in levels in proteins that had not been knocked out by CRISPR will imply that their interaction depends on the edited nodal protein.
LaCava predicts that these experiments will allow his team to create a biological story around the interaction of the deleted gene and all the other proteins that change in response to that.
“I felt that the team were responsive to my inquiries,” LaCava describes his experience with Synthego. “I certainly have positive impressions and I would relay those positive impressions as such.”
He adds that in the current competitive market, it is really easy for researchers to lose their money down the drain from subpar services or exhausting customer support.
“Nobody has time for that, so when somebody’s doing it right I’m definitely shouting it out,” he said.
Do you want to use CRISPR knockout cells in your research? If so, check out our free Cell Engineering 101 eBook.