Using site-directed mutagenesis, we generated two mutants that carry the D1849N (missense) and W1952* (nonsense) mutations (Fig. 4a).
Following mutagenesis, we generated flies with eyes that contain both wild-type tissue (marked red) and mutant tissue (marked white).
Using site-directed mutagenesis, we generated HA-tagged point mutants (Fig. S5A).
To avoid potential clinical caveats of cardiomyocytes differentiated from retroviral or lentiviral derived-hiPSC (e.g., insertional mutagenesis), we generated several non-viral, transgene-free hiPSC for cardiomyocyte differentiation using a three plasmid, seven-factor (SOKMNLT; SOX2, OCT4 (POU5F1), KLF4, MYC, NANOG, LIN28, and SV40L T antigen) EBNA-based episomal system [25].
Using PB-mediated germline mutagenesis, we generated mouse mutants, each of which carries a single PB transposon insertion (Ding et al., 2005) (S.D., X.W. and T.X., unpublished data).
Using N-ethyl N-nitrosourea mutagenesis, we generated and characterized a mouse model (named draggen), carrying the equivalent point mutation (c.1744A>G; p.I582V) to that found in the patient with periodic paralysis and myotonia.
By structure-guided mutagenesis, we have generated wt and polyA-expanded variants of RUNX2 with enhanced or reduced CC stability and found that the stability of the RUNX2 CC has a critical role in determining the subcellular localization, aggregation and toxicity of the protein.
Published online: March 6, 2008 By using gene-driven N-ethyl-N-nitrosourea (ENU) mutagenesis screening [5], we generated mice with distinct point mutations in Foxp2, backcrossed onto clean genomic backgrounds.
PCR products used for en passant mutagenesis were generated with polyacrylamide-gel electrophoresis-purified primers (Biomers, Ulm, Germany).
The SCRIB internal deletions and point mutagenesis were generated using PCR-based site-directed mutagenesis.
Site-directed mutagenesis was generated by PCR and the identities of individual clones were verified by sequencing.
