Conventional genome engineering with CRISPR–Cas9 creates double-strand breaks
(DSBs) that lead to undesirable byproducts and reduce product purity. Here we report an …

Efficient site-directed insertion of heterologous DNA into a genome remains an outstanding
challenge. Recombinases that can integrate kilobase-sized DNA constructs are difficult to …

CRISPR-associated transposases (CASTs) enable recombination-independent, multi-
kilobase DNA insertions at RNA-programmed genomic locations. However, the utility of type …

Canonical CRISPR-Cas systems utilize RNA-guided nucleases for targeted cleavage of
foreign nucleic acids, whereas some nuclease-deficient CRISPR-Cas complexes have been …

Cell engineering is commonly limited to the serial manipulation of a single gene or locus.
The recently discovered CRISPR-associated transposases (CASTs) could manipulate …

CRISPR-Cas nucleases are powerful tools for manipulating nucleic acids; however, targeted
insertion of DNA remains a challenge, as it requires host cell repair machinery. Here we …

Conventional CRISPR–Cas systems maintain genomic integrity by leveraging guide RNAs
for the nuclease-dependent degradation of mobile genetic elements, including plasmids and …

CRISPR-associated transposases (CASTs) direct DNA integration downstream of target
sites using the RNA-guided DNA binding activity of nuclease-deficient CRISPR-Cas …

The ability to edit the genome is essential for many state-of-the-art experimental paradigms.
Since DNA breaks stimulate repair, they can be exploited to target site-specific integration …

This protocol describes a method for CRISPR–Cas9-mediated genome editing that results in
scarless and marker-free integrations of DNA into Saccharomyces cerevisiae genomes …