Apath is offering an improved
version of the HCV replicon
While the
HCV replicon system represents a robust cell-based HCV RNA
replication system, the transformation efficiency (10-6)
of the originally described replicon limited its usefulness as a
genetic tool. Dr. Keril Blight isolated a series of replicons
that have a dramatically increased transformation efficiency.
Blazing Blight 1 (BB1) was described by Dr. Rice at the AASLD
meeting in Atlanta in April 2000. Blazing Blight 7 (BB7) is the
latest and best version of the HCV replicon being made available by
Apath. BB7 has a transformation efficiency of 10-1 (i.e.
10% of cells transfected with BB7 RNA display replicon-dependent
G418 resistance). This level of transformation efficiency is
approaching the level required for reverse genetic analyses.
Consistent with this, experiments have shown that the level of
replicon replication following transfection of BB7 into cells is
sufficiently robust to allow for measurement of first cycle RNA
replication.
(see
Science 290(5498):1972-1975;Dec 8, 2000)
Below is
shown the results of electroporation of RNA transcribed in vitro
from cDNA of Dr. Blight's replicons. In each panel G418 resistant
colonies are shown. Pol- is an NS5B null mutant replication
deficient control. As can be seen, Blazing Blight 7 has the highest
transformation efficiency.
