TALEN-Mediated Knock-in Genome Editing in Silkworm

Genome editing is a powerful tool for the analysis of targeted genes. Takuya Tsubota and colleagues from the National Agriculture and Food Research Organization in Japan previously found that a novel knock-in system, precise integration into target chromosome (PITCh), allows the integration of a donor vector with the hsp90 promoter and GFP into the silkworm biogenesis of lysosome-related organelles complex 1, subunit 2 gene.

Then the team evaluated whether the same technique can be used for the knock-in of other silkworm genes. The silkworm ku80 gene was selected as a target and was efficiently mutated using a pair of transcription activator-like effector nucleases (TALENs).

Microinjection of TALEN mRNAs mixed with the donor vector resulted in significant expression of the GFP marker in G0 larvae. GFP expression was detected in G1 individuals, suggesting that the integrated donor vector can be inherited in the next generation. It was further found to be inherited in the G2 generation.

The PITCh system offers a versatile tool for the knock-in of various genes, and could contribute to the further promotion of sericultural studies.

For more information on this study, read the article in Journal of Insect Biotechnology and Sericology.