Massively parallel single-amino-acid mutagenesis.


Abstract

Random mutagenesis methods only partially cover the mutational space and are constrained by DNA synthesis length limitations. Here we demonstrate programmed allelic series (PALS), a single-volume, site-directed mutagenesis approach using microarray-programmed oligonucleotides. We created libraries including nearly every missense mutation as singleton events for the yeast transcription factor Gal4 (99.9% coverage) and human tumor suppressor p53 (93.5%). PALS-based comprehensive missense mutational scans may aid structure-function studies, protein engineering, and the interpretation of variants identified by clinical sequencing.

Submission Details

ID: 7wvUDLZY3

Submitter: Shu-Ching Ou

Submission Date: July 18, 2018, 5:55 p.m.

Version: 1

Publication Details
Kitzman JO;Starita LM;Lo RS;Fields S;Shendure J,Nat Methods (2015) Massively parallel single-amino-acid mutagenesis. PMID:25559584
Additional Information

Sequence Assay Result Units