Evidence map›Paper›PMID 34921331›Full record

ArticleApplied microbiology and biotechnology2022

Development of a flow cytometry-based plating-free system for strain engineering in industrial fungi.

Yu-Jing Yang, Yin Liu, Dan-Dan Liu, Wen-Zhu Guo, Li-Xian Wang, Xing-Ji Wang, He-Xin Lv, Yang Yang, Qian Liu, Chao-Guang Tian

Abstract read
PubMed Publisher
In one paragraph

Article in Applied microbiology and biotechnology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.

0numbers the graph read from it
0cells of the map it votes in
10citing papers in PubMed
7.3field-weighted citation impact, top 3% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

10 citing papers in PubMed, 32 citations in OpenAlex.

  1. Article
  2. Article
  3. Article
  4. Review
  5. Comparative proteomics analysis ofFrontiers in microbiology · 2024
    Article
  6. Article
  7. Article
  8. Effect of new methionine-based ionic liquid on theIranian journal of microbiology · 2023
    Article
  9. Article
  10. Evaluation ofJournal of fungi (Basel, Switzerland) · 2022
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors at 2 institutions in 2 countries.

Yu-Jing Yang *College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
Yin Liu *College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
Dan-Dan LiuKey Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Wen-Zhu GuoKey Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Li-Xian WangNational Technology Innovation Center of Synthetic Biology, Tianjin, 300308, China.
Xing-Ji WangLongda Biotechnology Inc, 276400, Linyi, Shandong, China.
He-Xin LvCollege of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
Yang YangCollege of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
Qian LiuKey Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China. liu_q1@tib.cas.cn.ORCID http://orcid.org/0000-0001-6596-6402
Chao-Guang TianKey Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China. tian_cg@tib.cas.cn.
Chinese Academy of Sciences · CNTianjin University of Science and Technology · CN

Funding

National Major Science and Technology Projects of China 2018YFA0900500National Natural Science Foundation of China 31771386National Natural Science Foundation of China 31972878National Natural Science Foundation of China 31972879Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project TSBICIP-KJGG-006Youth Innovation Promotion Association of the Chinese Academy of Sciences 2019180
6 · The paper itself

Abstract

Recent technical advances regarding filamentous fungi have accelerated the engineering of fungal-based production and benefited basic science. However, challenges still remain and limit the speed of fungal applications. For example, high-throughput technologies tailored to filamentous fungi are not yet commonly available for genetic modification. The currently used fungal genetic manipulations are time-consuming and laborious. Here, we developed a flow cytometry-based plating-free system to directly screen and isolate the transformed protoplasts in industrial fungi Myceliophthora thermophila and Aspergillus niger. This system combines genetic engineering via the 2A peptide and the CRISPR-Cas9 system, strain screening by flow cytometry, and direct sorting of colonies for deep-well-plate incubation and phenotypic analysis while avoiding culturing transformed protoplasts in plates, colony picking, conidiation, and cultivation. As a proof of concept, we successfully applied this system to generate the glucoamylase-hyperproducing strains MtYM6 and AnLM3 in M. thermophila and A. niger, respectively. Notably, the protein secretion level and enzyme activities in MtYM6 were 17.3- and 25.1-fold higher than in the host strain. Overall, these findings suggest that the flow cytometry-based plating-free system can be a convenient and efficient tool for strain engineering in fungal biotechnology. We expect this system to facilitate improvements of filamentous fungal strains for industrial applications. KEY POINTS: • Development of a flow cytometry-based plating-free (FCPF) system is presented. • Application of FCPF system in M. thermophila and A. niger for glucoamylase platform. • Hyper-produced strains MtYM6 and AnLM3 for glucoamylase production are generated.

Indexed as

Gene EditingGlucan 1,4-alpha-GlucosidaseAspergillus nigerFlow CytometryGenetic EngineeringGlucan 1,4-alpha-GlucosidaseAspergillus nigerCRISPR-Cas9Flow cytometryGlucoamylaseMyceliophthora thermophilaPlating-free

Identifiers

PMID34921331
OpenAlexW4200231936

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.