TY - JOUR
AU - Karousis, Evangelos D.
AB -  Protein synthesis is a central process in gene expression and the development of efficient in vitro translation systems has been the focus of scientific efforts for decades. The production of translation-competent lysates originating from human cells or tissues remains challenging, mainly due to the variability of cell lysis conditions. Here we present a robust and fast method based on dual centrifugation that allows for detergent-free cell lysis under controlled mechanical forces. We optimized the lysate preparation to yield cytoplasm-enriched extracts from human cells that efficiently translate mRNAs in a cap-dependent as well as in an IRES-mediated way. Reduction of the phosphorylation state of eIF2α using recombinant GADD34 and 2-aminopurine considerably boosts the protein output, reinforcing the potential of this method to produce recombinant proteins from human lysates. 
TI - Production of human translation-competent lysates using dual centrifugation
JF - RNA Biology
DO - 10.1080/15476286.2021.2014695
DA - 2022-12-31
UR - https://www.deepdyve.com/lp/taylor-francis/production-of-human-translation-competent-lysates-using-dual-bwBneKPDE9
SP - 78
EP - 88
VL - 19
IS - 1
DP - DeepDyve
ER -