Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants.

Gemma F CodnerJoffrey MiannéAdam CaulderJorik LoefflerRachel FellRuairidh KingAlasdair J AllanMatthew MackenzieFran J PikeChristopher V McCabeSkevoulla ChristouSam JoynsonMarie HutchisonMichelle E StewartSaumya KumarMichelle M SimonLoranne AgiusQuentin M AnsteeKirill E VolynskiDimitri M KullmannSara WellsLydia Teboul

Published in: BMC biology (2018)

lssDNA donors reproducibly generate conditional alleles and can be used to introduce point mutations away from CRISPR/Cas9 cutting sites in mice. However, our work demonstrates that thorough quality control of new models is essential prior to reliably experimenting with mice generated by this method. These advances in genome editing techniques shift the challenge of mutagenesis from generation to the validation of new mutant models.

Keyphrases

crispr cas
genome editing
quality control
wild type
high fat diet induced
kidney transplantation
type diabetes
single molecule
binding protein
metabolic syndrome
nucleic acid