RGB marking with lentiviral vectors for multicolor clonal cell tracking.
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RGB marking with lentiviral vectors for multicolor clonal cell tracking. / Riecken, Kristoffer; Thomaschewski, Michael; Benten, Daniel; Fehse, Boris.
In: NAT PHYS, Vol. 7, No. 5, 5, 2012, p. 839-849.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - RGB marking with lentiviral vectors for multicolor clonal cell tracking.
AU - Riecken, Kristoffer
AU - Thomaschewski, Michael
AU - Benten, Daniel
AU - Fehse, Boris
PY - 2012
Y1 - 2012
N2 - Cells transduced with lentiviral vectors are individually marked by a highly characteristic pattern of insertion sites inherited by all their progeny. We have recently extended this principle of clonal cell marking by introducing the method of RGB marking, which makes use of the simultaneous transduction of target cells with three lentiviral gene ontology (LeGO) vectors encoding red, green or blue fluorescent proteins. In accordance with the additive color model, individual RGB-marked cells display a large variety of unique and highly specific colors. Color codes remain stable after cell division and can thus be used for clonal tracking in vivo and in vitro. Our protocol for efficient RGB marking is based on established methods of lentiviral vector production (3-4 d) and titration (3 d). The final RGB-marking step requires concurrent transduction with the three RGB vectors at equalized multiplicities of infection (1-12 h). The initial efficiency of RGB marking can be assessed after 2-4 d by flow cytometry and/or fluorescence microscopy.
AB - Cells transduced with lentiviral vectors are individually marked by a highly characteristic pattern of insertion sites inherited by all their progeny. We have recently extended this principle of clonal cell marking by introducing the method of RGB marking, which makes use of the simultaneous transduction of target cells with three lentiviral gene ontology (LeGO) vectors encoding red, green or blue fluorescent proteins. In accordance with the additive color model, individual RGB-marked cells display a large variety of unique and highly specific colors. Color codes remain stable after cell division and can thus be used for clonal tracking in vivo and in vitro. Our protocol for efficient RGB marking is based on established methods of lentiviral vector production (3-4 d) and titration (3 d). The final RGB-marking step requires concurrent transduction with the three RGB vectors at equalized multiplicities of infection (1-12 h). The initial efficiency of RGB marking can be assessed after 2-4 d by flow cytometry and/or fluorescence microscopy.
KW - Humans
KW - Flow Cytometry
KW - Microscopy, Fluorescence
KW - Transfection
KW - HEK293 Cells
KW - Genetic Vectors
KW - Cell Tracking/methods
KW - Green Fluorescent Proteins/analysis/genetics
KW - Lentivirus/genetics
KW - Luminescent Proteins/analysis/genetics
KW - Humans
KW - Flow Cytometry
KW - Microscopy, Fluorescence
KW - Transfection
KW - HEK293 Cells
KW - Genetic Vectors
KW - Cell Tracking/methods
KW - Green Fluorescent Proteins/analysis/genetics
KW - Lentivirus/genetics
KW - Luminescent Proteins/analysis/genetics
M3 - SCORING: Journal article
VL - 7
SP - 839
EP - 849
JO - NAT PHYS
JF - NAT PHYS
SN - 1745-2473
IS - 5
M1 - 5
ER -
