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Haploid Genetic Screens
in Human Cells
Unique experimental platform to identify
resistance mutations for cancer therapy
Haploid human cells are perfectly suited for genetic studies because the
inactivation of one gene copy is sufficient to elucidate its loss-of-function
phenotype. The obtained results are precise, robust and translate well into other
model systems.
Use Horizon’s haploid-based screening platform to elucidate the mechanism of
resistance of anti-cancer drugs, and use the knowledge of such resistance genes
to stratify patients and streamline clinical trials.
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Horizon Genomics GmbH, Campus Vienna Biocenter 3, 1030 Vienna, Austria
How do haploid genetic screens work?
1.
Haploid cells are mutagenized using retroviral gene trapping. This generates a population of cells in which
every cell line carries at least one gene trap insertions, thus resembling a gene knockout.
2.
Mutant pools are treated with a drug or drug candidate that affects cell viability or cell proliferation.
3.
Vast majority of mutants succumb to cell death, but a few drug resistant clones grow out.
4.
Location of gene trap insertions is determined by amplifying adjacent sequences by PCR, and next
generation sequencing.
Retroviral Gene
Trapping
Haploid Cells
Compound
Treatment
NGS
Gene Trap Mutants
Resistant Cells
Sequence
The ABT-737 screen in KBM-7 cells mutagenized with gene trapping
The screen resulted in two significant hits:
140
•
BAX: 51 independent insertions (p-value 10-111)
120
•
NOXA: 39 independent insertions (p-value 10-70)
Thus, results suggest important determinants for ABT-737
sensitivity and provide insights to the mechanism of action of
the drug.
100
-log (p-value)
Genetic inactivation of pro-apoptotic BAX or NOXA prevents
induction of apoptosis by ABT-737 and makes cells resistant
to the drug.
BAX (51 insertions)
NOXA (39 insertions)
80
60
40
20
0
0
100
200
300
400
500
600
700
800
Genes identified in screen (in alphabetical order)
Why to choose genetic screens in haploid cells?
UNBIASED
No previous assumption on mechanism of action
COMPREHENSIVE
Whole genome coverage, multiple gene trap insertions per gene
ROBUST
Unprecedented signal / noise ratio
PROVEN
Many high impact publications highlight potential
© Haploid Genetic Screens in Human Cells 2015 v-01
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Horizon Discovery, 7100 Cambridge Research Park, Waterbeach, Cambridge, CB25 9TL, United Kingdom