BibTex format
@article{Kopniczky:2020:10.1021/acssynbio.9b00437,
author = {Kopniczky, MB and Canavan, C and McClymont, DW and Crone, MA and Suckling, L and Goetzmann, B and Siciliano, V and MacDonald, JT and Jensen, K and Freemont, PS},
doi = {10.1021/acssynbio.9b00437},
journal = {ACS Synthetic Biology},
pages = {144--156},
title = {Cell-free protein synthesis as a prototyping platform for mammalian synthetic biology},
url = {http://dx.doi.org/10.1021/acssynbio.9b00437},
volume = {9},
year = {2020}
}
RIS format (EndNote, RefMan)
TY - JOUR
AB - The field of mammalian synthetic biology is expanding quickly, and technologies for engineering large synthetic gene circuits are increasingly accessible. However, for mammalian cell engineering, traditional tissue culture methods are slow and cumbersome, and are not suited for high-throughput characterization measurements. Here we have utilized mammalian cell-free protein synthesis (CFPS) assays using HeLa cell extracts and liquid handling automation as an alternative to tissue culture and flow cytometry-based measurements. Our CFPS assays take a few hours, and we have established optimized protocols for small-volume reactions using automated acoustic liquid handling technology. As a proof-of-concept, we characterized diverse types of genetic regulation in CFPS, including T7 constitutive promoter variants, internal ribosomal entry sites (IRES) constitutive translation-initiation sequence variants, CRISPR/dCas9-mediated transcription repression, and L7Ae-mediated translation repression. Our data shows simple regulatory elements for use in mammalian cells can be quickly prototyped in a CFPS model system.
AU - Kopniczky,MB
AU - Canavan,C
AU - McClymont,DW
AU - Crone,MA
AU - Suckling,L
AU - Goetzmann,B
AU - Siciliano,V
AU - MacDonald,JT
AU - Jensen,K
AU - Freemont,PS
DO - 10.1021/acssynbio.9b00437
EP - 156
PY - 2020///
SN - 2161-5063
SP - 144
TI - Cell-free protein synthesis as a prototyping platform for mammalian synthetic biology
T2 - ACS Synthetic Biology
UR - http://dx.doi.org/10.1021/acssynbio.9b00437
UR - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000508474400016&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR - https://pubs.acs.org/doi/10.1021/acssynbio.9b00437
UR - http://hdl.handle.net/10044/1/77915
VL - 9
ER -