Evidence map›Paper›PMID 36571722›Full record

ArticleCurrent protocols2022

NanoTag Nanobody Tools for Drosophila In Vitro and In Vivo Studies.

Ah-Ram Kim, Jun Xu, Ross Cheloha, Stephanie E Mohr, Jonathan Zirin, Hidde L Ploegh, Norbert Perrimon

Open access · greenAbstract read
In one paragraph

Article in Current protocols, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
1.2field-weighted citation impact, top 21% 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

8 citing papers in PubMed, 14 citations in OpenAlex.

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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

7 authors at 4 institutions in 2 countries.

Ah-Ram KimDepartment of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts.
Jun XuDepartment of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts.
Ross ChelohaBoston Children's Hospital and Harvard Medical School, Boston, Massachusetts.
Stephanie E MohrDepartment of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts.
Jonathan ZirinDepartment of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts.
Hidde L PloeghBoston Children's Hospital and Harvard Medical School, Boston, Massachusetts.
Norbert PerrimonDepartment of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts.
Boston VA Research Institute · USHarvard University · USBoston Children's Hospital · USChinese Academy of Sciences · CN

Funding

Functional genomics resources for the Drosophila - TR&D3P41GM132087 · NIGMS · HARVARD MEDICAL SCHOOL · PI PERRIMON, NORBERT · 2019 to 2023
$6.8M
NIGMS NIH HHS P41 GM132087
6 · The paper itself

Abstract

Nanobodies have emerged as powerful protein-binding tools to uncover protein functions. Using functionalized protein binders, proteins of interest can be visualized, degraded, delocalized, or post-translationally modified in vivo. We recently reported the use of two short peptide tags, 10-aa 127D01 and 14-aa VHH05, and their corresponding nanobodies, Nb127D01 and NbVHH05, for both in vitro and in vivo studies in Drosophila. Here, we provide detailed protocols for nanobody production and for visualization of proteins of interest in either fixed or live samples. In addition, we include protocols for endogenous protein tagging using CRISPR-mediated genome engineering. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Nanobody production in S2 cells Basic Protocol 2: Nanobody expression and purification in bacterial cells Basic Protocol 3: Immunostaining with nanobodies Basic Protocol 4: Immunoblotting with nanobodies Basic Protocol 5: Immunoprecipitation with nanobodies prepared from S2 cells Basic Protocol 6: Immunoprecipitation with nanobodies prepared from bacteria Basic Protocol 7: NbVHH05 and Nb127D01 used as chromobodies Basic Protocol 8: NanoTag trap as a method to alter protein localization Support Protocol: CRISPR-mediated tagging of endogenous genes with NanoTags.

Indexed as

Single-Domain AntibodiesAnimalsDrosophilaProtein BindingProtein TransportSingle-Domain AntibodiesCRISPRDrosophilaknock-innanobodyNanoTag

Identifiers

PMID36571722
PMCPMC9811555
OpenAlexW4313236544

What OpenQuestion holds

Textmetadata
LicenceTDM
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.