Evidence map›Paper›PMID 33661348›Full record

ArticleAnalytical and bioanalytical chemistry2021

Evaluation of CRISPR/Cas9 site-specific function and validation of sgRNA sequence by a Cas9/sgRNA-assisted reverse PCR technique.

Beibei Zhang, Jiamu Zhou, Miao Li, Yuanmeng Wei, Jiaojiao Wang, Yange Wang, Pingling Shi, Xiaoli Li, Zixu Huang, He Tang and 1 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Analytical and bioanalytical chemistry, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing 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

2 citing papers in PubMed.

  1. Article
  2. 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

11 authors.

Beibei ZhangHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Jiamu ZhouHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Miao LiHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Yuanmeng WeiHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Jiaojiao WangHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Yange WangHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Pingling ShiHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Xiaoli LiHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Zixu HuangHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
He TangHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China.
Zongming SongHenan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Henan Eye Institute, Henan Eye Hospital and Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, 450003, Henan, China. szmeye@163.com.

Funding

Science and Technology Department of Henan Province 192102310075
6 · The paper itself

Abstract

The effective application of the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system in biology, medicine and other fields is hindered by the off-target effects and loci-affinity of Cas9-sgRNA, especially at a genome-wide scale. In order to eliminate the occurrence of off-target effects and evaluate loci-affinity by CRISPR/Cas9 site-specific detection and screening of high-affinity sgRNA sequences, respectively, we develop a CRISPR/Cas9-assisted reverse PCR method for site-specific detection and sgRNA sequence validation. The detection method based on PCR can be used directly in the laboratory with PCR reaction conditions, without the need for an additional detection system, and the whole process of detection can be completed within 2 h. Therefore, it can be easily popularized with a PCR instrument. Finally, this method is fully verified by detecting multiple forms of site mutations and evaluating the affinity of a variety of sgRNA sequences for the CRISPR/Cas9 system. In sum, it provides an effective new analysis tool for CRISPR/Cas9 genome editing-related research. A CRISPR/Cas9-assisted reverse PCR method was developed for Cas9/sgRNA site-specific detection and sgRNA sequence validation. The technique detects target DNA in three steps: (1) target DNA is specifically cut by a pair of Cas9/sgRNA complexes; (2) the cleaved DNA is rapidly linked by T4 DNA ligase; (3) the ligated DNA is efficiently amplified by PCR (PCR or qPCR).

Indexed as

CRISPR-Cas SystemsBase SequenceClustered Regularly Interspaced Short Palindromic RepeatsDNADNA Mutational AnalysisGene EditingMutationPolymerase Chain ReactionRNA, Guide, CRISPR-Cas SystemsDNARNA, Guide, CRISPR-Cas SystemsCRISPR/Cas9sgRNA validationSite-specific detection

Identifiers

PMID33661348
PMCPMC7929911

What OpenQuestion holds

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Registered trials

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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.