RRC ID 22743
Author Sajwan S, Takasu Y, Tamura T, Uchino K, Sezutsu H, Zurovec M.
Title Efficient disruption of endogenous Bombyx gene by TAL effector nucleases.
Journal Insect Biochem Mol Biol
Abstract Engineered nucleases are proteins that are able to cleave DNA at specified sites in the genome. These proteins have recently been used for gene targeting in a number of organisms. We showed earlier that zinc finger nucleases (ZFNs) can be used for generating gene-specific mutations in Bombyx mori by an error-prone DNA repair process of non-homologous end joining (NHEJ). Here we test the utility of another type of chimeric nuclease based on bacterial TAL effector proteins in order to induce targeted mutations in silkworm DNA. We designed three TAL effector nucleases (TALENs) against the genomic locus BmBLOS2, previously targeted by ZFNs. All three TALENs were able to induce mutations in silkworm germline cells suggesting a higher success rate of this type of chimeric enzyme. The efficiency of two of the tested TALENs was slightly higher than of the successful ZFN used previously. Simple design, high frequency of candidate targeting sites and comparable efficiency of induction of NHEJ mutations make TALENs an important alternative to ZFNs.
Volume 43(1)
Pages 17-23
Published 2013-1-1
DOI 10.1016/j.ibmb.2012.10.011
PII S0965-1748(12)00162-2
PMID 23142190
MeSH Amino Acid Sequence Animals Base Sequence Bombyx / embryology Bombyx / genetics* DNA Mutational Analysis Deoxyribonucleases / metabolism Female Gene Targeting / methods* Genetic Vectors Male Molecular Sequence Data Open Reading Frames Saccharomyces cerevisiae
IF 3.827
Times Cited 52