RPROTK-RO

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The scaffolding function of LSD1/KDM1A reinforces a negative feedback loop to repress stem cell gene expression during primitive hematopoiesis.

Mattie J Casey et al.

iScience, 26(1), 105737-105737 (2023-01-04)

Lsd1/Kdm1a functions both as a histone demethylase enzyme and as a scaffold for assembling chromatin modifier and transcription factor complexes to regulate gene expression. The relative contributions of Lsd1's demethylase and scaffolding functions during embryogenesis are not known. Here, we

BioTAP-XL: Cross-linking/Tandem Affinity Purification to Study DNA Targets, RNA, and Protein Components of Chromatin-Associated Complexes.

Artyom A Alekseyenko et al.

Current protocols in molecular biology, 109, 21-21 (2015-01-07)

In order to understand how chromatin complexes function in the nucleus, it is important to obtain a comprehensive picture of their protein, DNA, and RNA components, as well as their mutual interactions. This unit presents a chromatin cross-linking approach (BioTAP-XL)

Limited time window for retinal gene therapy in a preclinical model of ciliopathy.

Poppy Datta et al.

Human molecular genetics, 29(14), 2337-2352 (2020-06-23)

Retinal degeneration is a common clinical feature of ciliopathies, a group of genetic diseases linked to ciliary dysfunction, and gene therapy is an attractive treatment option to prevent vision loss. Although the efficacy of retinal gene therapy is well established

Autolysis and inhibition of proteinase K, a subtilisin-related serine proteinase isolated from the fungus Tritirachium album Limber.

J Bajorath et al.

Biochimica et biophysica acta, 954(2), 176-182 (1988-05-18)

The activity of proteinase K (EC 3.4.21.14), a subtilisin-related serine proteinase, was assayed with azoalbumin that showed non-expected behavior in substrate saturation curve because of interaction between albumin molecules. Succinyl-(Ala)n-p-nitroanilide with n = 2 and 3, yielded specific activities of

PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins.

Jessica Spitzer et al.

Methods in enzymology, 539, 113-161 (2014-03-04)

We recently developed a protocol for the transcriptome-wide isolation of RNA recognition elements readily applicable to any protein or ribonucleoprotein complex directly contacting RNA (including RNA helicases, polymerases, or nucleases) expressed in cell culture models either naturally or ectopically (Hafner

How to Generate Non-Mosaic CRISPR/Cas9 Mediated Knock-In and Mutations in F0 Xenopus Through the Host-Transfer Technique.

Emmanuel Tadjuidje et al.

Methods in molecular biology (Clifton, N.J.), 1865, 105-117 (2018-08-29)

We have taken advantage of the well-established oocyte host transfer technique to optimize a method for CRISPR editing of Xenopus that provides an efficient non-mosaic targeted insertion of small DNA fragment through homology-directed repair mechanism.

IFN-γ-independent control of M. tuberculosis requires CD4 T cell-derived GM-CSF and activation of HIF-1α.

Erik Van Dis et al.

PLoS pathogens, 18(7), e1010721-e1010721 (2022-07-26)

The prevailing model of protective immunity to tuberculosis is that CD4 T cells produce the cytokine IFN-γ to activate bactericidal mechanisms in infected macrophages. Although IFN-γ-independent CD4 T cell based control of M. tuberculosis infection has been demonstrated in vivo

Comparative analysis and generation of a robust HIV-1 DNA quantification assay.

Jordan Thomas et al.

Journal of virological methods, 263, 24-31 (2018-10-17)

HIV-1 infection cannot be cured due to the presence of the latent reservoir (LR). Novel cure or treatment strategies, such as "shock and kill" or therapeutic vaccination, aim to reduce or eradicate the LR. Cure strategies utilise robust DNA quantification

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