Products/Services Used | Details | Operation |
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Custom DNA/RNA Oligos> | To express the MLH1dn protein, we generated an E. coli codon-optimized MLH1dn sequence (GenScript) and cloned it into the pET28a-His expression vector17. | Get A Quote |
Gene Synthesis> | Get A Quote | |
Synthetic sgRNA and crRNA Service> | Get A Quote |
The editing efficiencies of prime editing (PE) using ribonucleoprotein (RNP) and RNA delivery are not optimal due to the challenges in solid-phase synthesis of long PE guide RNA (pegRNA) (>125 nt). Here, we develop an efficient, rapid and cost-effective method for generating chemically modified pegRNA (125-145 nt) and engineered pegRNA (epegRNA) (170-190 nt). We use an optimized splint ligation approach and achieve approximately 90% production efficiency for these RNAs, referred to as L-pegRNA and L-epegRNA. L-epegRNA demonstrates enhanced editing efficiencies across various cell lines and human primary cells with improvements of up to more than tenfold when using RNP delivery and several hundredfold with RNA d... More