%0 Journal Article
%A Schönherr, Robert
%A Eichler, Nina
%A Sornaly, Fatama A.
%A Boger, Juliane
%A Frevert, Anne M.
%A Lahey-Rudolph, Janine Mia
%A Meyer, Hannah
%A Weymar, Lisa
%A Redecke, Lars
%T Intracellular protein crystallization in living insect cells
%J FEBS Open Bio
%V 15
%N 4
%@ 2211-5463
%C Hoboken, NJ
%I Wiley
%M PUBDB-2025-04364
%P 551 - 562
%D 2025
%X Crystallization of recombinant proteins in living cells is an emerging approach complementing conventional crystallization techniques. Homogeneous microcrystals well suited for serial diffraction experiments at X-ray free-electron lasers and synchrotron sources can be produced in a quasi-native environment, without the need for target protein purification. Several protein structures have already been solved; however, exploiting the full potential of this approach requires a systematic and versatile screening strategy for intracellular crystal growth. Recently, we published InCellCryst, a streamlined pipeline for producing microcrystals within living insect cells. Here, we present the detailed protocol, including optimized target gene expression using a baculovirus vector system, crystal formation, detection, and serial X-ray diffraction directly in the cells. The specific environment within the different cellular compartments acts as a screening parameter to maximize the probability of crystal growth. If successful, diffraction data can be collected 24 days after the start of target gene cloning.
%F PUB:(DE-HGF)16
%9 Journal Article
%R 10.1002/2211-5463.70020
%U https://bib-pubdb1.desy.de/record/639244