Individual Technology

Target Gene Clone Isolation Technology


Nozomu YACHIE
Research Center for Advanced Science and Technology, The University of Tokyo

theme

Abstract

The efficiency of any DNA assembly reaction is incomplete. Through genome editing, we have established a technology to selectively label and isolate only microbial cell clones having the desired reaction product after DNA assembly reactions.

Research Contents

Since the efficiency of any DNA assembly reaction is incomplete, the assembly reaction products require cloning through transduction into microbial cells and the evaluation by DNA sequencing. Consequently, samples obtained from reactions with lower efficiency require a larger number of clones for the evaluation, and this process becomes a bottleneck. We have established a selective clone isolation technology, which is as follows: after molecular DNA barcoding of assembly reaction products, the reaction product pool is analyzed all at once using the en masse sequencing technology, and then cells having the target reaction product are labeled and isolated in a DNA barcode-dependent manner by genome editing. This technology accelerates the throughput of the conventional DNA assembly process by 1000-fold.

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Strengths in the Industrial Arena

The construction ("Build" process) of long-chain DNA is the key component of the DBTL cycle. Without accelerating this process, the scale of useful microbial production processes cannot be expanded. Until recently, there was no method for efficiently obtaining the target DNA assembly product after DNA assembly. Due to this limitation, although efforts had been made to improve the efficiency of the DNA assembly itself, there was no method to substantially accelerate the "Build" process for various cases of DNA assembly. This technology resolves this situation.

References

Nishida K, Arazoe T, Yachie N, Banno S, Kakimoto M, Tabata M, Mochizuki M, Miyabe A, Araki M, Hara KY, Shimatani Z & Kondo A.: Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems, Science 353, aaf8729(2016)

Last updated:January 28, 2021