TL;DR
A practical protocol for using either PE or iPE to introduce point mutation(s) or small-to-medium sized insertions and deletions in cell culture, and how to assess editing efficiency via flow cytometry and next-generation sequencing is provided.
This study demonstrates donor-complementary prime editing (DoPE) as a one-step, DSB-free and library-compatible method for precise insertion of large DNA fragments without requiring recombinases or transposases.
CRISPR-Cas9 has transformed gene editing; however, it is based on doublestrand DNA breaks, which may result in unintentional mutations and structural alterations, thereby limiting its clinical translation. Instead of DSBs, new precision instruments, including prime editing, can be used to perform accurate search-an...
Recent advances in programmable large-DNA writing in the human genome are reviewed across four mechanistic categories: recombinase-, prime editing-, transposase-, and retrotransposon-mediated technologies.
A programmable gene replacement tool, named prime assembly (PA), which adapts prime editors to produce one or two pairs of 3'-flaps on both the genome and donor DNA, allowing megabase-scale genomic excision and/or kilobase-scale donor insertion at the gene of interest.
This chapter provides a practical protocol for base editing by electroporation by electroporation in adherent and suspension stem cells and details essential steps for cell handling and recovery, followed by standard readouts.
We use cookies to run the site and, with your consent, for analytics and to show ads. See our Cookie Policy.
