"Enhancement of transgene expression by mild hypothermia is promoter dependent in HEK293 cells"

Min Ho Jang*, Honggi Min*, Jae Seong Lee‡. Life 11(9): 901.

Mild hypothermia has been widely used to enhance transgene expression and improve the cellular productivity of mammalian cells. This study investigated mild hypothermia-responsive exogenous promoters in human embryonic kidney 293 (HEK293) cells using site-specific integration of various promoter seq..

"An optimized genome-wide, virus-free CRISPR screen for mammalian cells"

Kai Xiong*, Karen Julie la Cour Karottki, Hooman Hefzi, Songyuan Li, Lise Marie Grav, Shangzhong Li, Philipp Spahn, Jae Seong Lee, Ildze Ventina, Gyun Min Lee, Nathan E Lewis, Helene Faustrup Kildegaard, Lasse Ebdrup Pedersen‡. Cell Reports Methods 1: 4.

Pooled CRISPR screens have been widely applied to mammalian and other organisms to elucidate the interplay between genes and phenotypes of interest. The most popular method for delivering the CRISPR components into mammalian cells is lentivirus based. However, because lentivirus is not always an opt..

"A metabolic CRISPR-Cas9 screen in Chinese hamster ovary cells identifies glutamine-sensitive genes"

Karen Julie la Cour Karottki*, Hooman Hefzi, Songyuan Li, Lasse Ebdrup Pedersen, Philipp N Spahn, Chintan Joshi, David Ruckerbauer, Juan A Hernandez Bort, Alex Thomas, Jae Seong Lee, Nicole Borth, Gyun Min Lee, Helene Faustrup Kildegaard‡, Nathan E Lewis‡. Metabolic Engineering 66: 114-122.

Media and feed optimization have fueled many-fold improvements in mammalian biopharmaceutical production, but genome editing offers an emerging avenue for further enhancing cell metabolism and bioproduction. However, the complexity of metabolism, involving thousands of genes, makes it unclear which ..

"Streamlined human cell-based recombinase-mediated cassette exchange platform enables multigene expression for the production of therapeutic proteins"

Seunghyeon Shin, Su Hyun Kim, Jae Seong Lee‡, Gyun Min Lee‡. ACS Synthetic Biology 10(7): 1715-1727.

A platform, based on targeted integration of transgenes using recombinase-mediated cassette exchange (RMCE) coupled with CRISPR/Cas9, is increasingly being used for the development of mammalian cell lines that produce therapeutic proteins, because of reduced clonal variation and predictable transgen..

"Hydroxyurea selection for enhancement of homology-directed targeted integration of transgenes in CHO cells"

Jin Myeong Kwak*, Youngsik Lee*, Sung Wook Shin*, Jae Seong Lee‡. New Biotechnology 62: 26-31.

Site-specific integration via genome editing technologies has been implemented in Chinese hamster ovary (CHO) cells for predictable and efficient cell line development and engineering. Various strategies have been employed to enhance knock-in (KI) efficiency for precise homology-directed repair (HDR..

"Controlling ratios of plasmid-based double cut donor and CRISPR/Cas9 components to enhance targeted integration of transgenes in Chinese hamster ovary cells"

Sung Wook Shin*, Dongwoo Kim*, Jae Seong Lee. International journal of molecular sciences 22(5): 2407.

Chinese hamster ovary (CHO) cells are the most valuable expression host for the commercial production of biotherapeutics. Recent trends in recombinant CHO cell-line development have focused on the site-specific integration of transgenes encoding recombinant proteins over random integration. However,..