In a bid to discover how to maximize the tobacco plant's potentials as
bio-factories, an international team of scientists investigated
whichCRISPRtechnology was more efficient in incorporating two
herbicideresistance modifications simultaneously in the tobacco'/SuRB
gene/. The results not only exhibited which CRISPR technology was more
efficient, but that it was also possible to inherit gene-targeted
modifications.

Tobacco (/Nicotiana tabacum/) is a plant often considered for use as a
bio-factory in generating medicines and vaccines, while
theCRISPR-Casnuclease technology allows the induction of site-specific
double-strand breaks for better homologous recombination-mediated gene
targeting. Considering that gene targeting technology continues to be
enhanced by replacing SpCas9 with other CRISPR-Cas-based nucleases, the
scientists decided to test SaCas9 and a temperature-insensitive
ttLbCas12a using tobacco plant to target the/SuRB/gene using an
optimized protocol for Agrobacterium-mediated tobacco transformation and
tissue culture.

The scientists found that ttLbCas12a outperformed SaCas9 in terms of
gene targeting efficiencies by up to a third of the inoculated
cotyledons. They recorded that the conversion tract length of the gene
targeting reaction can stretch to over 250 bp long, and they were also
able to obtain multiple heritable heterozygous and non-T-DNA integrated
biallelic gene target events in tobacco. This is the first time that
researchers are able to obtain CRISPR-Cas-based heritable gene-targeted
events in tobacco.

Efficient gene targeting in Nicotiana tabacum using CRISPR/SaCas9 and
temperature tolerant LbCas12a - Huang - - Plant Biotechnology Journal -
Wiley Online Library [onlinelibrary.wiley.com]