CRISPR-Enabled Autonomous Transposable Element (CREATE) for RNA-based gene editing and delivery

Abstract

To realize the potential of genome editing for broad therapeutic applications, new tools are needed for delivery of multi-kilobase (kb) payloads at desired genomic sites with high precision. Here we introduce the CRISPR-Enabled Autonomous Transposable Element (CREATE), an RNA-based genome editing system that merges CRISPR/Cas9 with the human L1 retrotransposon to insert gene-sized payloads without DNA donors or double strand breaks. CREATE enables L1-mediated reverse transcription and integration of an RNA-encoded payload gene specifically at two Cas9-induced nick sites. CREATE is delivered using mRNA components and achieves integration of >1 kb payload in mammalian cells, opening the door to mRNA mediated therapeutic genome editing in vivo.

Allele Sails: launching traits and fates into wild populations with DNA sequence modifiers

“…As another point of reference, as of 2019, half of the pathogenic variants in the human ClinVar database were point-mutations, and almost 90% of clinically relevant insertions and deletions were less than 30bp 60 . Finally, recent work shows that larger fragments of DNA (which would albeit be considered transgenes) can be copy-pasted from one location to another using engineered retrotransposons 61,62 . These observations suggest it may soon be possible to push larger fragments of DNA into a population in a self-limiting manner using an editing locus transmitted in a Mendelian manner…”