This is the Gabbay Award lecture I presented at Brandeis University in Boston on Nov 16th 2009

1. ‘ Designer babies’: twenty years on Alan H Handyside London Bridge Fertility, Gynaecology And Genetics Centre London United Kingdom Gabbay Award Lecture Brandeis University November 16 th , 2009

4. Handyside et al Nature (1990) 344, 768 Pregnancies from biopsied human preimplantation embryos sexed by Y-specific amplification Born July, 1990 Gender identification and selective transfer of unaffected female embryos in X-linked disease Amplification of DYZ1 using 40 cycles of PCR from single cleavage stage blastomeres

6. Strategies for single cell genetic analysis SINGLE OR MULTIPLE CELL BIOPSY LYSIS NESTED PCR OUTER PCR MULTIPLEX F-PCR MULTIPLEX PCR PEP/DOP-PCR INNER PCR HETERODUPLEX FORMATION PAGE MDA AUTOMATED SEQUENCER SEQUENCING MINISEQUENCING AUTOMATED SEQUENCER SEQUENCING MINISEQUENCING AUTOMATED SEQUENCER MULTIPLEX PCR ANY COMBINATION OF CONVENTIONAL DNA BASED TESTS CGH MICROARRAYS A B C D E CGH DNA free reagents and conditions

7. Fiorentino et al (2005) Eur J Hum Genet 13, 953

8. Handyside et al (2004) Mol Hum Reprod 10, 167

9. Heterozygous Allele dropout Amp failure Allele specific preferential amplification Locus 1 cell 2 cells 5 cells Chromosome 7 CFTR exon 10 and intragenic marker (PCR and F-PCR) CF ΔF508 97/100 2 2 2 1 2 1 2 2 2 1 2 2 2 2 1 CF ΔF508 97/100 1 2 2 2 2 1 2 2 1 2 2 2 2 1 2 2 2 2 2 2 1 2 2 2 2 CF ΔF508 148/151 1 2 2 2 1 2 2 1 2 2 2 2 1 2 2 2 2 1 2 2 1 1 CF 17βTA 247/251 2 2 2 1 1 1 2 1 2 2 1 1 1 2 1 1 1 2 1 2 1 Chromosome 6 STR markers for HLA matching (F-PCR) D6S276 205/221 2 1 2 2 2 1 2 1 2 1 1 1 1 2 2 1 1 MIB 180/184 2 2 1 2 2 2 2 2 1 1 1 2 2 1 1 1 1 1 1 2 1 2 1 2 D6S291 157/163 2 2 2 1 2 2 1 2 2 1 1 2 1 2 1 1 1 2 1 1 HLABC 114/144 1 2 1 1 2 2 1 1 1 1 1 1 1 2 1 1 1 1 D6S265 113/121 1 1 1 1 2 2 1 1 2 2 1 2 2 1 2 1 1 2 1 1 2

10. Preimplantation genetic haplotyping following MDA from single cells Renwick et al (2006) Reprod Biomed Online 13, 110

11. Embryo 2 Monosomy 16, 21 Trisomy 22

13. CONTROL TEST

14. CONTROL TEST 1.0 1.5 0.5

15. CONTROL TEST 1.0 1.5 0.5

16. CONTROL TEST 1.0 1.5 0.5

17. First or first and second polar body biopsy on day 0 or day 1 Array comparative genomic hybridisation (CGH) following whole genome amplification

20. ♂ ♀ A A B B B A B A A A B A A A B A B A A A B A A B

21. B C D ♂ ♀ A Karyomapping combines genome wide linkage based detection of single gene defects (A) with chromosomal aneuploidy including monosomy/deletions (B) and trisomies involving inheritance of two different meiotic chromosomes from one parent (D). Chromosome duplication is not detected (C).

22. Family 1

23. Family 2 Preimplantation genetic diagnosis for cystic fibrosis Whole genome amplification by isothermal multiple displacement amplification of 2-10 cells in each biopsy/embryo

24. Embryo 7 13, 16, 18, 21, 22 Monosomy 18, 21, 22 X, Y, 21 Trisomy X Monosomy 21 Array CGH Karyomap

25. Mutation detection Multiplex PCR and minisequencing Preimplantation genetic haplotyping Fluorescence in situ hybridisation Array CGH Quantitative SNP array analysis and Karyomapping Single gene defects Single or combination  Any  Any Exclusion     SGD + HLA typing     Chromosome screening Aneuploidy   (5-12 chr)  (24 chr)  (24 chr) Meiotic aneuploidy  (PB only)  (PB only)  Trisomy only With parental origin     With meiotic origin     Mosaicism    Duplications/deletions    Uniparental disomy    Translocation chromosome imbalance Reciprocal/Robertsonian     Normal vs balanced     With 24 chr aneuploidy     Other Multifactorial recurrence risk  Copy Number Variants  Errors Allele dropout    Contamination   