Evidence map›Paper›PMID 40640356›Full record

ReviewNature protocols2025

Assembly and delivery of large DNA via chromosome elimination in yeast.

Yuan Ma, Yao Xiong, Jiayu Xu, Hui Xu, Zongheng Fu, Guang-Rong Zhao, Yi Wu, Ying-Jin Yuan

Abstract readReview
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In one paragraph

Review in Nature protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
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

4 citing papers in PubMed.

  1. Review
  2. Review
  3. Review
  4. Review
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

8 authors.

Yuan Ma *State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.ORCID 0009-0002-5040-0339
Yao Xiong *State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.
Jiayu Xu *State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.ORCID 0009-0007-9498-5208
Hui XuState Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.
Zongheng FuState Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.ORCID 0000-0002-0416-9482
Guang-Rong ZhaoState Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China.
Yi WuState Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China. yi.wu@tju.edu.cn.ORCID 0000-0002-2539-9228
Ying-Jin YuanState Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, China. yjyuan@tju.edu.cn.

Funding

National Key R&D Program of ChinaNational Natural Science Foundation of China (National Science Foundation of China) [32471483]Natural Science Foundation of TianjinNatural Science Foundation of Tianjin City (Natural Science Foundation of Tianjin) [23JCYBJC00220]
6 · The paper itself

Abstract

Manipulation of large-scale genetic information provides a powerful approach to deciphering and engineering complex biological functions. However, the manipulation of large DNA, such as assembly and delivery, remains complex and difficult. Here we describe the experimental design strategy and protocol for a chromosome elimination-mediated large DNA assembly and delivery method (HAnDy), which enables efficient Mb-scale DNA assembly and delivery in yeast conveniently. This protocol is divided into three parts: (1) CRISPR-Cas9-mediated elimination of chromosome, which includes design and integration of a synthetic single-guide RNA (sgRNA) site near the centromere, activation of chromosome elimination by mating, and verification of the chromosome elimination. It can be used to eliminate multiple chromosomes, achieving haploidization in yeast. (2) Haploidization-mediated DNA assembly, which includes the design and construction of initial assembly strains, DNA assembly by programmed haploidization and verification of the assembled clones. (3) Haploidization-mediated DNA delivery, which includes the design and construction of inducible haploidization strains, DNA delivery by programmed haploidization and verification of the delivered clones. Users can utilize this protocol entirely or selectively depending on their needs. With the use of this protocol, it takes 10 d to achieve chromosome elimination and 7-11 d to achieve a standard cycle of haploidization-mediated DNA assembly or delivery. This protocol provides an efficient approach that is useful for the elimination, assembly and delivery of large DNA in yeast, requiring basic molecular biology skills.

Indexed as

Chromosomes, FungalDNADNA, FungalGene EditingSaccharomyces cerevisiaeCRISPR-Cas SystemsHaploidyRNA, Guide, CRISPR-Cas SystemsDNADNA, FungalRNA, Guide, CRISPR-Cas Systems

Identifiers

What Socratic holds

Textmetadata
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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 Socratic graph.