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PCR Cloning and Subcloning> | … Fragment B containing floxed-miniSTOP (393 bps) was amplified from a plasmid that was synthesized by GenScript. The floxed-miniSTOP sequence is shown in Supplementary Materials. Fragments A, B, and C were combined using the Gibson Assembly Cloning Kit (#E510, … | Get A Quote |
Achromatopsia is characterized by amblyopia, photophobia, nystagmus, and color blindness. Previous animal models of achromatopsia have shown promising results using gene augmentation to restore cone function. However, the optimal therapeutic window to elicit recovery remains unknown. Here, we attempted two rounds of gene augmentation to generate recoverable mouse models of achromatopsia including a model with a knock-in stop cassette in intron 5 using -CRISPR (fficient dditions with sDNA nserts-CRISPR) and targeted embryonic stem (ES) cells. This model demonstrated that only 20% of CNGA3 levels in homozygotes derived from target ES cells remained, as compared to normal CNGA3 levels. Despite the low percentage ... More