Evidence map›Paper›PMID 42771154›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2027

Inducible Protein Knockdown via Engineered Small RNAs in Chlamydia.

Asha Densi, Ming Tan, Christine Sütterlin

Abstract read
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In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2027. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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

0 citing papers in PubMed.

No citing paper in PubMed yet.

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

3 authors.

Asha DensiDepartment of Developmental and Cell Biology, University of California, Irvine, CA, USA.
Ming TanDepartment of Microbiology and Molecular Genetics and Department of Medicine, University of California, Irvine, CA, USA.
Christine SütterlinDepartment of Developmental and Cell Biology, University of California, Irvine, CA, USA. suetterc@uci.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Genetic manipulation of Chlamydia trachomatis is challenging because of its obligate intracellular lifestyle and reduced genome with a high proportion of essential genes. Functional analysis of chlamydial genes often requires alternative genetic strategies such as conditional knockdown rather than gene disruption. This chapter describes the use of an engineered small RNA (sRNA) to achieve inducible protein knockdown in C. trachomatis. This approach utilizes the C. trachomatis sRNA CtrR3 as a scaffold to express a targeting sequence that interferes with a target mRNA by binding its 5' untranslated region (UTR), in the region of the ribosome-binding site (RBS) and start codon. The protocol detailed here covers design of the targeting sequence, bioinformatic analysis to minimize off-target binding, construction of the knockdown sRNA plasmid, generation of a chlamydial transformant expressing the engineered sRNA, induction of protein depletion, assessment of knockdown efficiency and phenotypes, and complementation to demonstrate knockdown specificity.

Indexed as

Bacterial ProteinsChlamydia trachomatisGene Knockdown TechniquesRNA, Small Untranslated5' Untranslated RegionsGenetic EngineeringHumansRNA, BacterialRNA, Small Interfering5' Untranslated RegionsBacterial ProteinsRNA, BacterialRNA, Small InterferingRNA, Small UntranslatedChlamydiaComplementationEngineered sRNAKnockdown specificityOff-target effectsPost-transcriptional repressionProtein depletionProtein knockdown

Identifiers

PMID42771154

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.