Allan Bradley

TL;DR: Investigation of coelectroporation as a method for introducing minor genetic changes into specific genes in embryonic stem cells found that 3 of 36 of the 6-TG-resistant clones had the desired 4-bp insertion without any other disruption of the HPRT locus.

TL;DR: Two cell lines that can be routinely transmitted through the germline of chimeras after cloning and prolonged selection in tissue culture are developed and a variety of methods for generating non-selected mutations at the X-linked hprt locus in ES cells are established.

TL;DR: The principles underpinning genetic screens in mouse ES cells with examples of previously successful screens are described and protocols are provided for piggyBac transposon-mediated mutagenesis, production of the corresponding homozygous mutants in a Blm-deficient genetic background, and methods for mapping and validation of mutations recovered from screens of such libraries.

TL;DR: This method can provide homozygous mammalian loss-of-function mutants for forward genetic applications by constructing DNA vectors that are selectable based on their copy number and using the piggyBac transposon to limit the initial mutagenesis to one copy per cell.

TL;DR: In this article, a desired non-human animal or an animal cell or human cell which contains a predefined, specific and desired alteration in at least one of its two p53 chromosomal alleles, such that one of these alleles contains a mutation which alters the expression of the allele, and the other of the alleles expresses either a normal p53 gene product, or comprises an identical or different p53 mutation.