Generation of BAC Transgenic Epithelial Organoids

TY - JOUR

T1 - Generation of BAC Transgenic Epithelial Organoids

AU - Schwank, G.

AU - Andersson-Rolf, A.

AU - Koo, B.K.

AU - Sasaki, N.

AU - Clevers, H.

N1 - Reporting year: 2013

PY - 2013

Y1 - 2013

N2 - Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of these ex vivo cultures is their accessibility to live imaging. So far the establishment of transgenic fluorescent reporter organoids has required the generation of transgenic mice, a laborious and time-consuming process, which cannot be extended to human cultures. Here we present a transfection protocol that enables the generation of recombinant mouse and human reporter organoids using BAC (bacterial artificial chromosome) technology.

AB - Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of these ex vivo cultures is their accessibility to live imaging. So far the establishment of transgenic fluorescent reporter organoids has required the generation of transgenic mice, a laborious and time-consuming process, which cannot be extended to human cultures. Here we present a transfection protocol that enables the generation of recombinant mouse and human reporter organoids using BAC (bacterial artificial chromosome) technology.

U2 - 10.1371/journal.pone.0076871

DO - 10.1371/journal.pone.0076871

M3 - Article

SN - 1932-6203

VL - 8

SP - 76871

JO - PLoS One

JF - PLoS One

IS - 10

ER -