TY  - JOUR
AU  - Schönherr, Robert
AU  - Eichler, Nina
AU  - Sornaly, Fatama A.
AU  - Boger, Juliane
AU  - Frevert, Anne M.
AU  - Lahey-Rudolph, Janine Mia
AU  - Meyer, Hannah
AU  - Weymar, Lisa
AU  - Redecke, Lars
TI  - Intracellular protein crystallization in living insect cells
JO  - FEBS Open Bio
VL  - 15
IS  - 4
SN  - 2211-5463
CY  - Hoboken, NJ
PB  - Wiley
M1  - PUBDB-2025-04364
SP  - 551 - 562
PY  - 2025
AB  - 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.
LB  - PUB:(DE-HGF)16
DO  - DOI:10.1002/2211-5463.70020
UR  - https://bib-pubdb1.desy.de/record/639244
ER  -