• Multiple sgRNA: Quadruple firepower, precise strikes

The cleavage efficiency of a single sgRNA is often significantly reduced due to poor target accessibility and chromatin status. A study published in Nature Communications in 2026 proposed a simple and effective solution: using four sgRNAs to simultaneously target one gene.

Multiple sgRNAs operate through a dual mechanism of redundancy and synergy: even if one fails, the others can still be replaced, and large fragment deletions frequently occur between cleavage sites, significantly increasing the proportion of complete gene inactivation. The study evaluated the activity of over 2,000 sgRNAs in 33% of the Drosophila genome, achieving targeting efficiency exceeding 99%, an off-target rate below 1%, outperforming the Cas9 system in parallel comparisons with over 100 genes, and identifying phenotypes missed by traditional methods, providing an efficient and reliable new tool for large-scale functional genomics screening.

• Chemically modified sgRNA: Fully armed with guide RNA

Natural sgRNA is easily degraded by nuclease enzymes in the body, severely limiting the application of CRISPR in vivo.

The research team systematically introduced various chemical modifications to obtain gRNA with over 90% nucleotide modification. Although some modification patterns exhibit sequence-dependent activity differences, the team has established an expanding library of highly modified gRNAs that maintain or even enhance editing efficiency both in vitro and in vivo, clearing key bottlenecks for in vivo delivery and therapeutic applications of CRISPR.

GentleGen has built a gene service technology platform integrating automation and intelligence, embedding an integrated link between gene synthesis, oligonucleotide synthesis, high-throughput sequencing, and gene editing services, providing a truly standardized, highly precise, and cost-effective one-stop solution for CRISPR sgRNA library construction and screening.

Relying on its self-developed oligonucleotide synthesis platform, GentleGen has achieved an ultimate mutation rate of 1‰ in oligonucleotide synthesis—meaning that for critical sequence lengths, GentleGen can ensure that over 90.5% of sgRNA sequences perfectly match theoretical designs, guaranteeing precise targeting of every gRNA from the sequence source!

Coverage and homogeneity are the "golden dual cores" for evaluating document quality. Under a strict quality control system, GentleGen delivers over 99% library coverage to clients, while maintaining an industry-leading level of uniformity below 10.

• High-standard quality control + NGS deep sequencing verification, data speaks for itself at a glance

Once the library is built, it is not simply a matter of packaging and shipping. GentleGen independently validates every delivered plasmid library strictly according to NGS sequencing depth ≥300×, delivering high-purity, endotoxin-free plasmids (total ≥ 200μg), allowing you to control quality during the "first mile" of experiments without extra effort.

There are many directions for basic research or drug development. For whole-genome exploration, GentleGen can directly provide standardized human and mouse whole-genome knockout libraries to support large-scale genetic screening efforts. For specific needs for precisely targeting specific signaling pathways or target gene sets, GentleGen's customized services are always on standby, tailoring and synthesizing each client's unique targets.

• From screening to validation, chain-based delivery enables closed loops to arrive faster

Relying on strong NGS bioanalytical capabilities and long-term gene editing experience, GentleGen is not limited to library plasmid construction—from lentiviral packaging, cell infection, screening pressure application, to final high-throughput sequencing and data analysis, GentleGen can scientifically and reasonably integrate generations, helping researchers shorten project cycles, quickly lock onto "targets," and achieve a complete validation closed loop.

GentleGen offers a one-stop sgRNA library service with high precision, high efficiency, and high cost-performance, defined precisely by standards! GentleGen can provide related services. For more details about related services, please send an email to marketing@gentlegen.com.