THE CHALLENGE
The major challenge in developing reliable gene editing solutions for livestock lies in delivering CRISPR reagents efficiently into mammalian zygotes without harming embryo viability, an issue that significantly limits scalability, consistency, and commercial viability. Traditional methods like microinjection are technically challenging, time-consuming, and often damage embryos, while common alternatives such as electroporation, lipofection, or viral delivery face barriers like poor uptake, low editing precision, and biosafety concerns. These inefficiencies result in low biallelic (both copies of a gene) knockout rates, high mosaicism (uneven editing across cells), and hidden genetic effects due to lingering maternal RNA. For agri-biotech companies, this translates to higher costs, longer development cycles, and inconsistent outcomes in breeding programs, ultimately slowing down the path from innovation to market. Addressing these hurdles with a more reliable and scalable delivery platform is essential to unlock the full commercial and scientific potential of gene editing in livestock.
OUR SOLUTION
CRISPR-DART offers a breakthrough in livestock gene editing by using two precisely timed electroporation rounds to efficiently deliver gene-editing CRISPR-cas ribonucleoproteins into bovine zygotes. This method enables highly reliable, over 90% biallelic deletion of the targeted DNA, significantly reducing mosaicism and avoiding the need for complex microinjection or multiple costly procedures. By combining DNA editing with RNA targeting in one streamlined step, it also eliminates maternal mRNAs that can hide true genetic effects in cleavage embryos. For agri-biotech companies, this translates into faster, more predictable results, improved embryo survival, and scalable, cost-effective production of genetically enhanced cattle for traits like disease resistance or productivity—making CRISPR-DART a powerful platform for next-generation animal breeding and research.
Figure: Schematic of CRISPR-DART procedure. Created with BioRender.com.
Advantages:
- >90% biallelic deletion efficiency
- Simultaneous DNA and RNA targeting (CRISPR-DART)
- Large deletions (>1 kb) for complete gene disruption
- Minimal or no impact on embryo survival
Potential Application:
- Heat-tolerant and hornless cattle breeding
- Gene-edited livestock for biomedical disease models
- Xenotransplantation-ready pigs for organ donation
- Transgenic animals for biopharmaceutical protein production
