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The shift from traditional oligo synthesis to oligo pool synthesis technology signifies a profound transformation in gene synthesis techniques. This transition has carved out unprecedented and expansive prospects for various field of research, including gene editing, drug development, disease diagnosis, and disease treatment.
Oligo pools are comprised of a vast array of oligonucleotide sequences, all synchronously synthesized on microarray chips utilizing specialized synthesis technologies. Subsequently, these sequences are eluted and dissolved within individual microtubes. Oligo pools have applications in NGS hybrid capture probe libraries, CRISPR sgRNA libraries, and mutation libraries, thereby facilitating subsequent high-throughput screening endeavors.
Tradition vs. Innovation: Oligo Pools
Traditional oligo synthesis methods, due to its high cost, long cycle, and low efficiency, is gradually evolving into a major obstacle in the scientific research process. However, the emergence of oligo pool synthesis technology has completely broken this deadlock. By synthesizing a large number of oligonucleotides with different sequences at the same time, we can achieve high-throughput, low-cost, rapid, and efficient gene sequence preparation – providing unprecedented convenience and flexibility for scientific researchers.
Strategies for High-Throughput Oligo Pool Synthesis
In the field of oligo synthesis, two primary technologies dominate: semiconductor chip-based synthesis and inkjet synthesis. Each technology possesses their own unique advantages and are suited for different scenarios and requirements.
Semiconductor Chip Technology
Technical Features
- Develops molecular chips utilizing semiconductor technologies such as integrated circuits.
- Achieves high-density synthesis throughput, reducing the cost per base pair.
- Highly automated, enabling full-process automation management from sequence analysis and sequence optimization to synthesis and detection.
Advantages
- High-Throughput and Low-Cost: Offers significant advantages for high-throughput experiments requiring large-scale, low-cost primer synthesis.
- Precise Control: Ensures the quality and stability of oligos by precisely controlling conditions like temperature, humidity, and lighting during the synthesis process.
Inkjet Technology
Technical Features
- Utilizes inkjet technology to print DNA sequences point-by-point onto semiconductor chips.
- Precisely controls the liquid volume at each synthesis point, avoiding overuse and waste that are common in traditional batch synthesis methods.
- Further enhances raw material utilization and synthesis efficiency through optimization of synthesis parameters and conditions.
Advantages
- Customization and High Flexibility: Customizable oligo length, sequence, and modifications based on experimental requirements, suitable for rapid and flexible small-batch oligo synthesis.
- Low Equipment Cost: Compared to semiconductor chip synthesis, inkjet synthesis technology boasts a lower equipment cost.
The Many Applications of Oligo Pools
- sgRNA Gene Editing Libraries: Construct libraries for precise screening of optimal sgRNA sequences.
- shRNA Libraries: Rapidly build libraries to filter out effective targets, ensuring the accuracy of library construction.
- NGS Hybridization Capture Probe Libraries: Applicable for both capturing and detecting target genes, as well as screening specific probes for target sequences.
- FISH Probe Synthesis: High-throughput synthesis of probes with diverse sequences, covering multiple sites within the target region, enhancing the sensitivity and accuracy of FISH detection.
- Mutant Library Construction: Create mutant libraries, synthesizing diverse DNA regulatory regions to pinpoint those meeting research goals.
Downstream Application Case of Oligo Pools
LacIs (lactose repressor proteins) serves as a pivotal regulatory component of the lactose operon, yet it exhibits a certain degree of “leakiness” in its expression under natural conditions. Thus, limiting its utility in applications requiring precise expression control.
Synbio Technologies’ oligo pool synthesis service has greatly facilitated the authors of this case in successfully constructing multiple mutant libraries. Through high-throughput screening, they identified several LacI mutants that significantly reduced leakiness in their expression, with the M7 mutant standing out particularly. Leveraging the capabilities of the M7 mutant, the researchers constructed a high-value sugar biosensor and proposed a scheme to enhance galactosylation activity through biosensor-based screening.
Engineering and Application of LacI Mutants with Stringent Expressions
Synbio Technologies: Your Trusted Partner for Oligo Pool Synthesis
As a leader in oligo pool synthesis, Synbio Technologies is dedicated to providing customers with the highest quality oligo pools synthesis services in the industry. Their comprehensive Syno-HT synthesis platform uses inkjet technology to simultaneously synthesize tens of thousands of oligos on micro-semiconductor chips. This process ensures the guaranteed delivery of the precise sequences required by the customer, thus improving the efficiency and success rate of subsequent high-throughput screenings. Whether you need a CRISPR sgRNA screening library, high-throughput sequencing, or high-throughput gene synthesis, our customized services are the perfect match for your experimental needs and accelerating your R&D pipelines!
References
[1] Wu J, Liang C, Li Y, Zeng Y, Sun X, Jiang P, Chen W, Xiong D, Jin JM, Tang SY. Engineering and application of LacI mutants with stringent expressions. Microb Biotechnol. 2024 Mar;17(3):e14427.
[2] Uzonyi A, Nir R, Schwartz S. Cloning of DNA oligo pools for in vitro expression. STAR Protoc. 2022 Jan 17;3(1):101103.
[3] Kuiper BP, Prins RC, Billerbeck S. Oligo Pools as an Affordable Source of Synthetic DNA for Cost-Effective Library Construction in Protein- and Metabolic Pathway Engineering. Chembiochem. 2022 Apr 5;23(7):e202100507.
