Engineered TnpB genome editors for plants and human cells identified by ribonucleoprotein mutational scanning
Brittney W. Thornton·David F. Savage·Cynthia I. Terrace·Rachel F. Weissman·Julia Tartaglia·Brenda Duong·Ugrappa Nagalakshmi·Viktoriya Georgieva·Savithramma P Dinesh-Kumar·Myeong-Je Cho·Evan D. Groover·Ryan V. Tran·George Austin·Jennifer A. Doudna·Jorge E. Rodriguez·Flora Zhiqi Wang·Jung-Un Park
TnpB is a diverse family of RNA-guided endonucleases associated with prokaryotic transposons. Because of their small size and putative evolutionary relationship to CRISPR-Cas12, TnpB enzymes hold great potential for genome editing. However, most TnpBs lack robust gene-editing activity. Here, we mapped comprehensive sequence-function landscapes of a TnpB ribonucleoprotein using deep mutational scanning and we discovered activating mutations in both the RNA and the protein. Leveraging the protein'
