We have almost 40 different CRISPR knockin/out projects at different stages of completion. Twenty projects have progressed to the point that live founder mice with the desired allele have been generated. Most of these projects are point mutation knockins, one project is an indel gene knockout, two are deletions (1 and 16kb). The percentage of mice with the desired knockin allele has averaged 25%, ranging from 2-88% of the mice delivered. This is 4 founder mice carrying the knockin allele per project on average, ranging from 1-10 founder mice per project. Where we have deep sequencing data on the mutant knockin allele, 70% of the founder mice have the desired mutant allele in more than 45% of the reads. Given the prevalence of the mutant allele in founder mice, it is not surprising that in projects that have progressed to the point of mating, all have transmitted the mutant allele to the next generation.
ISTT SURVEY ON PRODUCTION OF TRANSGENIC MICE
Thomas J. Fielder, Laura Barrios, Lluís Montoliu
PCA Graph 1
"This is a two-dimensional graph that was made by projecting the mean of the
individual facility in relation to Principal Components 1 and 2.
In this graph, the X-axis (corresponding to Principal Component 1) is interpreted
as “Transgenesis efficiency” or “Transgenic pups obtained”. Facilities located to
the right have better values than those located to the left. The Y-axis
(corresponding to Principal Component 2) is interpreted as “embryo transfer
efficiency” or “Pups born”. Facilities located above zero have better values than
those located below. The ideal position for a facility is in the upper right
quadrant. Facilities showing the best performance regarding the ratios
considered will be found in this upper right corner.
Your facility is depicted as an upside down blue triangle while the other
facilities are depicted as green open circles.
According to the position of your facility, the following recommendations should
be considered. If your facility is located in the upper right quadrant, don’t
change anything. If your facility is located in the upper left quadrant, total pup
yields are good, but you may be able to improve your transgenic pup yields. If
your facility is located in the lower right quadrant, transgenic pup yields are
good, but you may be able to improve your total pup yields (e.g., by improving
the embryo-transfer procedure). Finally, if your facility is located in the lower left
quadrant, you may be able to improve both total pup and transgenic pup yields."