End-Modified Double-Strand DNA Donors for Clone-Free & Selection-Free Gene Knock-In in Mammalian Cells Lines

Dr. Zhao from the University of IL has developed a rapid, cloning-free CRISPR/Cas9 based gene knock-in (KI) method that utilizes Cas9/gRNA ribonucleoprotein( Cas9 RNP) and end-modified double-strand DNA (dsDNA) generated by PCR. This method allows large gene (up to 1.1kb) knock-in with at least 5% KI efficiency in various mammalian cell lines. With high KI efficiency and short preparation time, this method can be applied in mammalian genome editing and engineering.