Key references for human germline genetic editing Narrative outline of our project: Human Germline Genetic Modification Reviews Nucleic acids delivery methods for genome editing in zygotes and embryos Recent (2016) review of the methods available - see Table 1 and Figure 1 for an overview of methods. Limited consideration of CRISPR and/or sperm selection to enhance efficiency. Refs 67-74 in that paper are also relevant for our project. In vivo Gene Transfer into Testis and Sperm: Developments and Future Application Another good review, it references e.g. In vivo transfection of testicular germ cells and testis-mediated gene transfer which succeeded in generating transgenic sperm. Gene transfer to sperm and testis also does not consider the impact of CRISPR or cell sorting and only deems methods viable if the mice can produce transgenic progeny from natural mating (no IVF), but is otherwise a good review. Cell Sorting Single Cell Isolation and Analysis Summary of cell sorting techniques (FACS/MACS/microfluidics), specific example here: A method for high purity sorting of rare cell subsets applied to TDC. Allows us to extract transgenic sperm even if they are a tiny fraction of the total. Sperm selection using magnetic activated cell sortinq has been investigated as well. CRISPR Efficient precise knockin with a double cut HDR donor after CRISPR/Cas9-mediated double-stranded DNA cleavage Much-improved method for CRISPR-based gene knock-in, another related technique is MMEJ-assisted gene knock-in. Both allow the insertion of transgenes at specific locations in the genome. Base Editing Correction of the Marfan Syndrome Pathogenic FBN1 Mutation by Base Editing in Human Cells and Heterozygous Embryos Base editing in human embryos; has limited utility for now but it's a good option for single point mutations. Preimplantation Genetic Diagnosis (PGD) 1 EFTA00804855