DETECTION
Fluorescence imaging shines a light on systems biology Chromosomal tagging using advanced fluorescent labeling techniques can help us to understand cells at a systemic level. Researchers in Germany have developed a universal approach for CRISPR-Cas12a-assisted PCR tagging of mammalian genes, using scalable live cell imaging to optimize their methods. The Knop Lab at the University of
a Knop Lab PhD student working in
enables the rapid creation of
Heidelberg’s Center for Molecular
collaboration with the German Cancer
mammalian cell lines with targeted
Biology is researching the processes
Research Center, explained: “Our group
large chromosomal insertions, such as
that regulate cell signaling, cell
focuses on answering questions about
green fluorescent protein (GFP). These
differentiation, protein quality control
how cells work, and obtaining a
proteins can then be visualized with a
and homeostasis in yeast and in
systemic understanding of them. The
microscope or cell imaging system. The
clinically-relevant mammalian systems.
aim is to develop tools and methods for
aim is to establish a universal, scalable
The group combines live cell imaging
use by the wider research community,
and high-fidelity method to tag
with a range of fluorescence-based
to answer biological questions and
proteins in mammalian cells, enabling
techniques to observe protein function
develop a better understanding about
researchers to tag and target as many
and investigate biomolecular
cells’ behavior.”
proteins as possible.”
environment. These functional
“My focus is on method development,”
The group’s PCR tagging method
high-content imaging methods are
Krisztina continued. “Currently, I am
involves the transfection of a gene-
combined with genetic and genomic
working on optimizing a PCR tagging
specific PCR cassette into the target
approaches to explore the cellular
method, a one-step procedure for
cells, together with a helper plasmid
processes of interest. Krisztina Gubicz,
chromosomal gene tagging that
containing a Cas12a endonuclease,
interactions in their natural
The Knop Lab team (NOTE: Photo taken with respect to social distancing measures and COVID regulations within Germany.) 24
TECAN JOURNAL 2/2021
