lentiCRISPR v2

CRISPR Mouse - Repression CCR-5

Experiment
CRISPR Mouse - Repression CCR-5
Product
lentiCRISPR v2 from Addgene
Manufacturer
Addgene

Protocol tips

Protocol tips
There is no need to perform a negative control golden-gate reaction (without
insert) as it will always contain colonies so not a good indicator of cloning success.

Publication protocol

Plasmids, antibodies and cell lines
pLentiCRISPR.v2, pAdeasy-1, pShuttle-CMV and pLentiLox3.7 (pLL3.7) were from Addgene. pNL4-3 BaL + (R5-tropic, containing the backbone of NL4-3 and the Env of BaL, referred to as BaL hereafter) and pNL4-3 (X4-tropic), as well as TZM-bl and C8166-CCR5 cell lines were from NIH AIDS Research & Reference Reagent Program, Division of AIDS, NIH. pCH042, a transmitted/founder HIV-1 molecular clone were kindly provided by Professor Beatrice H. Hahn ( Parrish et al., 2013 ). Anti-CCR5-PE (clone 2D7) and matched isotype control antibodies were from BD Pharmingen. Rabbit anti-Cas9 polyclonal antibody was purchased from Hangzhou HuaAn Biotechnology, and the internal control antibody (anti-β-actin) was purchased from Santa Cruz. CHO-CCR5, a CHO-K1 cell line with stable CCR5 expression was described previously ( Hu et al., 2005 ).

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Reviews

lentiCRISPR v2 from Addgene has not yet been reviewed for this experiment

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Discussion

Discussion

4 years ago

Author: Mario Udinese Italy

Floxing mice with CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Discussion

5 years ago

Author: Ben Saar Israel

How to choose a region to target for CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Mouse - Repression CCR-5 using lentiCRISPR v2 from Addgene.

Paper title
Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9.
Full paper
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Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing CRISPR Mouse - Repression CCR-5 using lentiCRISPR v2 from Addgene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

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